Professional Documents
Culture Documents
UniverSoul Circus V Dallas
UniverSoul Circus V Dallas
DALLAS COUNTY
8/19/2015 12:55:03 PM
FELICIA PITRE
DISTRICT CLERK
DC-15-09369
CAUSENO. ____________- -
Christi Underwood
Plaintiff,
DALLAS COUNTY, TEXAS
v.
THE CITY OF DALLAS and DALLAS COUNTY
HEALTH AND HUMAN SERVICES, a Munjcipal
- - - JUDICIAL DISTRICT
Corporation,
Defendants.
Page 1 of 13
II. PARTIES
1.
Plaintiff SCI is a corporation organized under the laws of the State of Georgia
with a principal place of business at 230 Peachtree Street, N.W., Suite 2000, Atlanta, Georgia
30303.
2.
State of Texas and may be served with process by serving the Dallas City Secretary's Office
Attn: Rosa A. Rios, 1500 Marilla Street, Room 5D South, Dallas, TX 75201-6390.
3.
Texas, where the City is located and may be served with process by serving the Dallas City
Secretary's Office Attn: Rosa A. Rios, 1500 Marilla Street, Room 5D South, Dallas, TX 752016390.
Jurisdiction is proper in this Court because the amount in controversy meets the
minimum jurisdictional limits of this Court and all parties are subject to personal jurisdiction in
Texas.
5.
Venue is proper in Dallas County pursuant to Tex. Civ. Prac. & Rem. Code
15.002, because all or a substantial part of the events giving rise to this claim occurred in Dallas
County.
SCI, a nationally renowned circus, hereby brings this special proceeding pursuant
to the Tex. R. eiv. Pro. 680 and Tex. Civ. Prac. & Rem. Code, 65, and challenges the
Page 2 of 13
Defendants' decision to deny SCI a permit to exhibit two elephants, Betty and Bo, at SCI's
upcoming performances in Dallas, Texas.
SCI and its agents are duly and properly licensed by the federal government, as
7.
well as many state governments, including Texas, to exhibit elephants and other exotic animals.
SCI has been holding circus performances featuring elephants for the past 20
8.
years. SCI contracts with various vendors to provide its elephants and other exotic animals.
SCI's elephant vendor routinely tests its elephants for tuberculosis, complies with the most
recent Guidelines for the Control of Tuberculosis in Elephants, and has never exhibited an
elephant that poses a public health risk.
Defendants allege that Bo and Betty cannot be exhibited because they tested
9.
"reactive" to tuberculosis.
10.
and in fact is wholly arbitrary. In the event Defendants are relying on the DPP Vet TB Assay for
Elephants (the ''DPP Test") from December 2014, this test serves only as a screening test to aid
in the diagnosis of active tuberculosis. The DPP Test is not definitive and often gives false
positives. In fact, the only USDA-recommended diagnostic test to detect TB in live elephants
that is considered to be definitive is a mycobacterial culture of trunk: wash samples (the "Trunk
Wash Test"). See the supporting materials for the use of the Chembio Diagnostic Systems, Inc.
DPP Vet TB Assay for Elephants (the "DPP Test Supporting Materials") attached hereto as
Exhibit 1,
~3,
and the Affidavit of Dr. Michael D. Stafford (the "Stafford Affidavit") attached
Page 3 of 13
11.
Following the DPP Test in December 2014, which indicated that Bo and Betty
were Hreactive" to certain antibodies, the elephants underwent the more extensive and definitive
Trunk Wash Test on January 2,3, and 5,2015. The results of the Trunk Wash Test indicated that
Bo and Betty are not infected with or carriers of tuberculosis and that their December 2014 DPP
Test results were in fact false positives. See Stafford Affidavit, ~12.
12.
Betty and Bo underwent another Trunk Wash Test on June 12, 14, and 15, 2015.
The results of this Trunk Wash Test indicate that Betty and Bo are not infected with or carriers of
tuberculosis. See Stafford Affidavit,
~12
and also see the June 2015 Trunk Wash Test attached
Bo had Trunk Wash Tests performed every three months in 2011 and every six months
thereafter. Each Trunk Wash Test result has consi.stently been negative for tuberculosis. See
Stafford Affidavit,
~M!8-9,
When SCI shared the aforementioned information with Defendants and asked
Defendants to reconsider the decision to deny permits for exhibition of Bo and Betty, Defendants
maintained their prior decision. See a copy of an E-mail from Sarah Ausherman attached hereto
as Exhibit 3.
15.
SCI now brings this proceeding to challenge Defendants' position that SCI cannot
exhibit the two elephants that tested reactive to tuberculosis, despite definitive and
uncontroverted scientific and medical testing and analysis that definitively establishes the DPP
Test was a false positive and the elephants do not have tuberculosis. SCI hereby requests that
Defendants' denial of the permit to exhibit Bo and Betty be enjoined vacated or annulled on the
Page 4 of 13
grounds that such Defendants' denial of such permit was arbitrary and capricious, an abuse of
discretion, and/or contrary to applicable governing law, and issue an injunction requiring the
City to issue a permit for Bo and Betty to perform.
V. RELEVANT FACTS
16.
Since its inception, SCI has been in the business of promoting and performing
Throughout its history, some of SCI's most popular acts have involved the use of
elephants.
18.
SCI is scheduled to promote and perfonn its circus from August 19, 2015 through
August 30, 2015 in Dallas, Texas. Each of SCI's scheduled performances inv01ve the elephants
Betty and Bo, and SCI has advertised and promoted the exhibition of Betty and Bo.
19.
Despite
State of Texas, SCI received an email from the City on July 27,2015, stating that the City denied
permission for SCI to exhibit Bo and Betty. Attached to the email was a letter written by Zachary
Thompson, Director of DCHHS.
recommended that the City deny SCI permission to exhibit the elephants because they tested
reactive for tuberculosis in the December 2014 DPP Test. A copy of this communication is
attached hereto as Exhibit 4.
20.
In response to the aforementioned email, SCI sent the City additional information
regarding the two elephants, including information evidencing that the elephants do not have
tuberculosis and that the DPP Test was a false positive.
attached hereto as Exhibit 5.
Page 5 of 13
21.
The City forwarded the information to DCHHS, who reviewed the information
but maintained their earlier recommendation to the City, who denied SCI permission to exhibit
Bo and Betty. A copy of the letter from DCHHS regarding reconsideration is attached hereto as
Exhibit 6.
22.
DCHHS explained that allowing the exhibition of elephants that tested "reactive"
to tuberculosis posed a threat of potential trarurnission of tuberculosis from the elephants to the
public. The fear that any elephant that has ever tested "reactive" to tuberculosis poses a threat to
the public is misguided.
23.
DCHHS did not provide any scientific explanations for its decision, nor did its
agents speak with any large animal veterinarians or experts with experience in tuberculosis
testing, diagnosis, or transmission in elephants. Upon information and belief, the City and
DCHHS failed to consider the 2008 Guidelines for the Control of Tuberculosis in Elephants (the
"2008 Guidelines") or the 2010 Guidelines for the Control of Tuberculosis in Elephants (the
"2010 Guidelines"). Instead, DCHHS ignored the best scientific and medical evidence without
any basis.
24.
On August 17, 2015, a representative of SCI, Alan M. Briskin, sent a letter to the
City explaining that SCI followed the standards of care, testing, diagnosis, and treatment of Bo
and Betty as set forth in both the 2008 Guidelines and the 2010 Guidelines. See the letter from
Alan M. Briskin to the City which is attached hereto as Exhibit 7.
25.
The aforementioned letter from Alan M. Briskin also evidenced that the 2010
Guidelines are more stringent than the 2008 Guidelines, and that SCI's elephant vendor and, of
course, 80 and Betty, met the standards of care, testing, diagnosis, and treatment as set forth in
the 2010 Guidelines.
Page 6 of 13
26.
The 2010 Guidelines provide that elephants testing reactive or positive to a DPP
Test, but also having a negative culture from a trunk wash and no known exposure to TB fall
under Group 3C. The 2010 Guidelines prescribe how often elephants that fall under Group 3C
should be tested. The 2010 Guidelines specifically state that elephants should be "[mJonitor[ed]
by culture (triple sample method) every 3 months for one year, then every 6 months for life."
I
The 2010 Guidelines further state that elephants should undergo "repeat MAPINDPP testing
every 3 months for the first year, then every 6 months for an additional 2 years if elephant
remains . . . reactive. If all cultures remain negative after 3 years annual serological testing may
resume as described in these guidelines." The 2010 Guidelines then go further to state that "the
elephant may be able to travel and have public contact after 6 months of successful documented
USDA approved treatment." See a copy of the 2010 Guidelines attached hereto as Exhibit 8.
27.
In accordance with the 2010 Guidelines, Bo and Betty were tested. by Trunk. Wash
Tests every three months for one year, and have been tested by Trunk Wash Test every six
months thereafter. All Trunk Wash Tests have come back negative. Bo and Betty also repeated
the MAPIAlDPP testing
28.
Based upon the foregoing, Bo and Betty meet all of the requirements of exhibition
for the United States Department of Agriculture (''USDA''), the Animal and Plant Health
Inspection Service ("APHIS"), the United States Animal Health Association ("USAHA "), and
the State of Texas. See Stafford Affidavit,
29.
~13.
Additionally, Betty and Bo's testing and care was provided and supervised by
Michael D. Stafford, D.V.M., who fully complied with the most restrictive protocols related to
the elephants. See Stafford Affidavit.
Page 7 of 13
30.
Despite the overwhe1ming evidence that Bo and Betty do not have tuberculosis
and do not pose a threat to public health, Defendants' refused to reconsider their position.
Defendants provided no scientific evidence or expert opinion to explain their refusal.
SCI repeats and realleges all factual allegations as if fully set forth herein.
32.
Despite the clear language of the 2008 Guidelines and the 2010 Guidelines
regarding the screening role of DPP Testing and the definitive nature of Trunk Wash Testing,
Defendants have denied SCI pennission to exhibit Bo and Betty at SCI's Dallas, Texas
performances.
33.
SCI adhered to the standards set forth in the 2008 and 2010 Guidelines for caring,
testing, diagnosing, and treating tuberculosis in elephants. Additionally, SCI adhered to the 2008
and 20 10 Guidelines when it detennined that the elephants did not have tuberculosis and were
not a public health risk.
34.
Despite SCI's compliance with the 2008 Guidelines and the 2010 Guidelines,
Defendants have denied SCI permission to exhibit Bo and Betty at SCI's Dallas, Texas
performances.
35.
SCI denies that there are any other standards upon which it is required to adhere
regarding the control, care, testing, diagnosis, and treatment of tuberculosis in elephants.
36.
Pursuant to Tex. Civ. Prac. & Rem Code 37, et. Sec., SCI is entitled to an order
declaring that SCI adhered to the 2008 Guidelines and the 2010 Guidelines, thus Defendants are
required to permit the exhibition of Betty and Bo at SCI's upcoming performances in Dallas,
Texas.
Page 8 of 13
37.
Pursuant to Tex. R . Civ Pro. 37.009 Plaintiff seeks the recovery of its reasonable
SCI repeats and realleges all factuaJ a11egations as if fully set forth herein.
39.
SCI seeks injunctive relief under both equitable and statutory principles, Tex.
R.Civ. Pro. 680-693, and Tex. Civ. Prac. & and Rem.Code 65.011.
40.
Defendants have refused to permit SCI to exhibit Betty and Bo at SCI's upcoming
performances in Dallas, Texas, although Betty and Bo do not have tuberculosis and are not a
public health risk.
41.
Defendants have not provided any explanation for their decision, except for their
unfounded fear that the elephants are a public health risk. Defendants have refused to consider
any evidence to the contrary.
acts and omissions, and SCI will continue to be irreparably harmed if it is W1able to exhibit Betty
and Bo as part of its 2015 performances in Dallas, Texas.
43.
SCI has no adequate remedy at law and the calculation of damages that may be
Prior to commencing this proceeding, SCI has not sought injunctive or similar
relief for its claims against Defendants in this Court or any other court.
45.
Betty and Bo, andlor restraining Defendants from unlawfully interfering with SCI's exhibition of
Page 9 of 13
elephants as long as SCI continues to comply with all applicable rules and regulations governing
the exhibition of the elephants, including the 2010 Guidelines.
46.
without notice to Defendants, and immediate injunctive relief be awarded that will restrain and
enjoin Defendants and their applicable agents, servants, employees, attorneys and those persons
in active concert with them as follows:
a.
Enjoining Defendants from prohibiting the exhibition of elephants Betty and Bo;
b.
From interfering with the lawful exhibition of elephants Betty and Bo;
c.
From interfering with or prohibiting the display or exhibition within the City of
Dallas, circus elephants who have been tested for elephant tuberculosis by the Trunk Wash Test
in accordance with USDA, USAHA and APHIS guidelines and whose test results were negative;
d.
respect any documents, computer files (including, but not limited to, emails, hard drives, disc
drives, zip drives), communications, data d.rafts
Of
permitting or the denial of permitting for SCI to exhibit elephants in the City of Dallas.
WHEREFORE, SCI demands judgment as follows:
1.
On the first cause of action, declaring that a circus which abided by the 2008 and
2010 Guidelines for the Control of Tuberculosis in Elephants and whose elephants tested
negative for tuberculosis in the Trunk Wash Test is entitled to exhibit the elephants in the City of
Dallas;
2.
On the second cause of action, enjoining Defendants from interfering with SCI's
Of,
Defendants to permit SCI to exhibit Betty and Bo upon the condition that SCI continues to
Page 100f13
comply with all applicable rules and regulations pertain1ng to the care and maintenance of
tuberculosis in elephants, as set forth in the 2010 Guidelines;
3.
On all counts, the costs of this suit and reasonable attorneys' fees; and
4.
For such other and further legal and equitable relief as this Court deems just and
proper.
RespectfuUy submitted:
FERGUSON, BRASWELL & FRASER,
By:
P.e.
Page 11 of 13
Pursuant to Dallas LR 2.02 The undersigned counsel has provided opposing counsel at
least two (2) hours' notice before this Application for TRO and accompanying proposed order
were presented to the Court for decision.
To the best of the undersigned's knowledge, the case in which this application for TRO is
presented is not subject to transfer under Local Rule 1.06
Page 12 of13
VERIFICATION
SlATE OF GEORGIA
)
)
COUNTY OF FULTON
I, Cedric Walker, the PresUient of Soul Circus, Inc., being first duly sworn, verifies that the allegations
set forth in thls Original Petition and Application for Temporary Restraining Ottler and T~pora.cy and
Permanem Injunctions, ex.cepr those made upon infonnatioo and belief, are, withio my personal knowledge
and to the best of my knowledge and belief,
By.
Cedric Walke!
Its:
Presidetlt
truE:
and accurate.
CATALOG 1/ 6!-9110-0
2 Te&t Kit
HEMBIO
DIAGNOSTIC
~nrEMs,
INC.
INTENDED USE
PRINCIPLE OF TEST
The DPP VetTa Assay is a single use Immunochromatographic rapid test for the detection of IIntibodies to
M,ycobacterjwn tJlberculo.r/.! and Mycobacterium bovl.s In
serwn, plasma or whole blood from African elephants
(Loxodonta alr/eana) and Asian elophllllts (Elephas
nUTXlmus) . Tho test is used as an aid in the diagnosis of act! ve
tub~rculosls (TB) in conjunction with othot diagnostic
method8,
The Chembio DPP YetTB Aasay is bll.Sed 00 immunochromatographic technology. The test employs two
recombinant antigens, an M. fl<bercu/w/s and an M bovls
antigen, whloh ate separately immobilized on lhe membrane
solid phase. rt alBo utilizes recombinant Proteln AlO conjugated
to colloidal gold plll1lcles for antibody deteGtion. The DPP
VerrB Assay uscs ~erum, plasma or whole blood. The sample
Is applied to tho SAMPLE+BUPPER well with the buff6r.
After thc sample and buffer have migrated onto the test strip
additional buffer 15 added 10 the BUFFER well. Thc buffer
facJUtates the latoral flow of tho released products aod
promotes the binding of antibodies to the antigens. If present,
the antibodies bind to the gold conjugllted antibody binding
protein, in a reactive sample, the dye cor\.iugated-immune
complex migrates on the nltroc~llulose ml:lllbrane and is
captured by the antigens immobilized in the TBST (1 2) area
producing 8 plnkJpurple line. In tho absonco of detectable
antibody, no specific immune complex would be fonned on the
lest llna. and, therefore, no pink/pw-p\e line would appear In Ihe
TEST (I 2) area. Unbound conjugated gold particles continue
to migrate olong the membrane and produce a pink/purple lin"
in the CONTROL (C) ar~. This proudursl control serves to
demonstrate lhat the reagents have been properly applied and
have migrated through the device.
MATERIALS PROVIDED
Each kit contains the following iterru:
2 DPP VetTB test devices
Product lnsert
Disposable gloves
Exhibit 1
10-6294-0 Rev 2
Nov.mb..,. 2012
i>"oS" I of4
TEST PROCEDURE
I.
2,
PRECAUTIONS
I.
2.
3.
4.
S.
3.
Figure 1
Teal WIndow wllh
BUFFER--+nJ!:-'
Well 2
7.
8.
._~~t-
SAMPLE
+ BUFFER
WeUl
4.
5.
6.
SPECIMEN COLLECTION
The DPP VetTH Assay can be
serum or plasma.
p~lformed
on whole blood,
IO~294 0 R.av 2
Novcrnber 20 12
PaRe 2 of4
two
~r"l1~~~~
30D C.
6.
7.
8.
Nonreactive Rerult
Only a pink/purplo CONTROL (C) line is visible. The sample
contains no detectable BI1Iibody to bolh TB and
mycobacteriosis antigens. A nonreactive result does not
preclude the possibility ofTB infection,
NO n,ST (I 2) Line,
QUALITY CONTROL
A pink/purple colored line should Blwa)'s appear in
CONTROL (C) area if the tesl has been perfonned correctly
and the device Is working properly. It serves as an Internal
test procedlll'al control.
INTERPRETATION OF RESULTS
Iuv!lIid Result
A pink/purple Bne should always appear In the CONTROL (C)
area, whether Dr not II line appears in the TEST area. If there is
no distinct pink/purple line In the CONTROL (C) area., the test
is invalid and should be repeated using a new devtee,
Reactive Result
I , Three pink/purple lines. one line In the CONTROL area,
one line in the TEST (1) area Ilnd ono line In the TEST (2)
area indicates a reactive result. Thl" suggests that the
satnple is reactive fur TB.
No CONTROL (el Li ,.
LIne
line are
for TB.
vlsibl~.
th~
sample is reactive
Do not open the ~ealed fall pouch until Just prior to use.
Do not use kit contents beyond labeled expiration date,
7.
Read r~ults in a well-lit are!!..
8. A reactive result using tbl': Chembio DPP VetTD AlIsay
suggests the presence of antibodIes to M. ,uberr:u/O.f/J andJor
M boy/so The Chemblo DPP VetTS Assay is intended as sn
aid In th~ diagnosis and treatment of TB in elephants.
9. For a reactive result, the intensity of tho to.!! IInc does nol
necessaril y correlate wi lh the titer of antibody in the
specimen.
10. A non-reactive result does not preclude tne pass ibliity of
cxposure to TB or lnfectlon with TB. An antibody response
to recent CXP08UI'e may take several mootbs to reach
detectable levels.
II . In treated elephants, interpret rellults with caution:
a. Treatment against TB may reduce antibody responses,
thus resultIng In non-reactive results in some cases.
5.
6.
I().62940 Rov 2
Novembor 1012
PlSO j of4
PERFORMANCE CHARACTERISTICS
732.
3.
Sensitivity
4.
Specificity
lli1d
Table 1.
Diagnostic seusltlvU), 01 DPP VerrB AJny
DPP VetTe
Mycobacterial
species isolated
M. tuberculosis
M bovls
M. I1.Iberculo.Yls
Elephant
species
AITicen
Aftlcan
Asian
reactive
6/6
III
33/33
ORDERING INFORMATION
Cat II
65-9110-0
65-9111-0
Tobie 2.
~pecies
non-reactive
79179
68/68
African
Asian
65-9112-0
1'nInk wash
culture negative
79179
611/68
I'./.TI ""
uh,ll.,j
."\lIlIoll'
REPRODUCIBILITY STUDIES
Reproducibility was evaluatoo at three independent
laboratories using two serials of DPP VetTB Assay. A
roference pane! of 40 blindly-coded $8mples representing
negatl ve, weakly reactive and strongly react! ve sera wel'C
tested 3 times on 3 different days . The complied results fi'om
3 laboratories demomrtraled 98.6% accuracy.
REFERENCES
I.
2.
10-62940 Rov 2
November 20 \ 2
Page 4 of4
Product
DPPYctTB 2 Test Kit
DPPYetTB 5 Test Kit
DPPYetTB 20 Test Kit
Antigens licensed from
Statens Serum Institut
www.ssi.dk
CAUSE NO . _ _ _ _ _ _ __
Plaintiff,
DALLAS COUNTY, TEXAS
v.
THE CITY OF DALLAS and DALLAS COUNTY
HEALTH AND HUMAN SERVICES, a Municipal
Corporation,
Defendants .
regarding the matters set forth herein. I am a veterinarian, and I have personal knowledge of the
facts stated in this Affidavit. This Affidavit is given volWltarily.
2.
I am licensed to practice in the State of Missouri, and have been practicing since
3.
I have been the veterinarian for Larry Carden's elephants, Betty and Bo, since 2011
1989.
and have reviewed all their records and diagnostic testing back to 2008.
4.
Both Betty and Bo have been tested using the Trunk Wash test and Stat-Pale test
since 2008.
5.
The Trunk Wash test is a diagnostic test used to determine whether an elephant has
tuberculosis by taking three samples from the elephant's trunk on three separate days.
Page 1 of 3
ExHIBIT
2
.0IIII
6.
The
Stat~Pak test
The
Stat~Pak
test became unavailable in 2012 and we started using the DPP Vet TB
Assay for Elephants (the "DPP Test"), which was approved as the replacement to the
Stat~Pak
test
in 2013.
8.
Stat~PaklDPP
was reactive in 2009). As such, we followed the procedures set forth in the 20 10 Guidelines for
treatment. That is, we tested both Bo and Betty every three months using both the Trunk Wash
Test and DPPlMAPlA tests for the first year, then every six months thereafter.
9.
Each Trunk Wash Test came back negative. The DPP tests came back reactive as
well as the MAPIA tests; however, the MAPIA tests improved after Bo and Betty were treated
using a three-drug cocktail suggested by the 2008 and 2010 Guidelines.
10.
In 2013 and 2014 the DPP tesis from ChemBio were very scarce, which did not
The OPP Test serves only as a screening test to aid in the diagnosis of active
tuberculosis. The DPP Test is not definitive and often gives false positives. The only
USDA~
recommended defmitive diagnostic test to detect TB in live elephants is the Trunk Wash Test.
12.
Following the DPP Test in December 2014, which found that Bo and Betty were
"reactive" for tuberculosis, as always, the elephants underwent the more extensive and defmitive
Trunk Wash Test on January 2,3, and 5, 2015. The results of the Trunk. Wash Test indicated that
Bo and Betty do not have tuberculosis and their December 2014 DPP Test results were false
positives, which means that they only showed antibodies were present.
Page 2 of 3
13.
B6 and Betty ~urr6Iltly meet all of the reqUirements for exhibition prt'lscrlbed by
the United States Depart:b1ent of Agriculture (UUSDA ")t the Animal fl!ld Plant Health Inspection
Service ("APHIS"), the United States Anitn.al Health Association ("OSAHA'1, and the State of
Texa.s. Moreover. Dr. Sidwa. the Trow State Public :Health Veterinarian, who is in charge of
approviiIg whether an animal is safe fur traval in the
travel and agreed that thoy both met the 2010 Guidelines.
~A~~
.
.
.
/~1D. Stafford
Sworn to and sUbset:1bed
. .
bofote me this
~ ,
lq- day
of AuguS"t 2015
Notary Public
My Commission'E~les:
Pego 3 of3
EXHIBIT A
Date ofTests
Bo
Betty
Result
Negative
X
X
X
Negative
Negative
Negative
Negative
2010
December 20,21,22,2010
2011
--_.
March 17, l8, 19, 20!1
June 10,11, 12, 2011
August 18, 19,20,2011
December 13,14, is, 2011
2012
May 7, 8,9,2012
May 23,24, 25,2012
August 22, 23, 24, 2012
December 18,19, 20,2012
2013
June 25,26,27,2013
December 11, 12, 13,2013
X
X
Negative
Negative
Negative
NegatIve
X
X
Negative
Negative
X
X
X
X
Negative
Negative
Negative
2014
June 16, 17,18, 2014
Janua ry 2, 3, 5, 2015
i 2015
June 12, 14, 1S, 2015
"-
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12/29/10
ONE LARGE SMOOTH COLONY RECOVERED FROM
BI PLATE
01120/2011
fout units of me6iwn are inoculated for
Aoid fast
weaks,
Default
~ulture
result iB
06 NEGA~rV8
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COLTURB
JENNY GRAHAM
SUPERVISOR
MYCOBACTERIOLOGY LAB
CARDEN CIRCUS,BO
CAP (1.21789-01
2PAGE
END OF REPORT
Advanced Diagnostic Laboratories
~ATIONAL JEWISH HEALTH
1400 Jackson Street
DENVER, CO 80206
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1 PAGE
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90206'
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DATE
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END OF REPO~T
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3PAGE
END OF REPORT
DENVeR, co 80206
clien t Servi ces] 800-5 50-62 27
CAP #2'1789-01
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LAB 108 /S,OR\
4. 2011 II:07AM
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JENNY GRAHAM
SUPERVISOR
MYC08ACTERIOLOGY LAB
TRUNK WASI{
03/17{ 11
DENN1S SCf{MIDl'
mo
COMMENTS
SMEAR BXAMINATION
No
AFB CULTURE
3/24/2011
CULTQRE NEGATtVE FOR MYCOBACrERIA
W2698
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04/30/ 11
PArrENT'S PHYSICIAN
CARDEN C!RCUS,BETTY
'.\~~lNY GI~AHAM
TRUNK WASH
SUPERVISOR
03118/11
DENNIS SCHMITT
CONTINUli:D
Adv~ced Diagn ostic Labo ratori es
NATIONAL
l~OO
J~W!SH
HRALTH
Jacks on stree t
DENVER, CO 80206
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03/19/11
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6/16/2011
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06/11/11
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1 PAGE
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cUnr~flr.b!@nic.ocg
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6/15/2011
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06/12/11
DE..tomtS SCHM':TT
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6/16/2011
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BO
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CULTUF3
3/3
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NO, 46! 6
if positiv e,
3PAGE
END OF REPORT
~dvnnced
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CAP #2178901
80206
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12
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II
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I
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06/10/11
DENNIS SCMITT
417 861 9572
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611612011
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FOR MYCOBACTERlll
07/22/11
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SOURCE
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05/11/H
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61, 833 9141
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12
NO. 4615
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06n2/11
PENNIS SCHMITT
417 861 9572
PHYSICIAN'S ?AX NO
SMEAR BXAM!NAT!ON
No AFB found
CARDEN CIRCOS/BETTY
6116/2011
2PAOE
CONTINUED
Advanced Diagnostic Laboratories
NATIONAL JEWISH
1400
HEAL~H
J~ckson St~eet
DENVER/ CO 80206
Client. Services: 800-550-6227
CAP #21789-01
ellA (106005807
)..AB lOS (8.08)
NO. 4615
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NATIONAL
Labo~atoriea
~EWlgH
HEALTH
90206
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SPRINGFIELD,MO 65803
T9496
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08/18/2Q~1
08:00
SPECIMEN SOURCE
ACTUAL COLLECT DATE
PATIENT'S PHYSICIAN
?HYSIC~&~'S PHONE
~HYSICIAN'S FAX NO
SMEAR EXAMItIlF\T:;:ON
TRUNK Wl\SH
APB CO!.rTURE
APB CULTURE COMMENT
PENDING
Four units of meiurn are inoculate~ fo r
recovery of the or~anisro, Acid fast
bacilli (AFB) ~re ,ead at 1/3 / and 6
weeks , Default culture result i9
"PE:NDING' and should be int~rpreted
as NEGATIVE until ~ positLve result
is en t ered or ~hB final neg~t i ve
result is entere~ at n waaks. Eight
week LJ culture I"ill be reported
only if positive.
T949B
COL~:
SPECIMEN SOURCE
Ar.rOAL COLLECT DATE
S PH'(SICIAN
PHYSICIAN'S PHONE
PHYSICIAN'S FAX NO
CARDEN
DBNNIS
SCHMI'r ~1
(NOAfEI
08/18/11
CIRC~S,BO
TRUNK WMH
08 / 19 / 11
DK,l\INIS SCHi'lITT
417 861 9572
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80206
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n 2011
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,V\,w.NJI.nbM/&
TEST RESULTS
SMEAR
ADx
5/6
?,
T949B
NO, 6225
REC:
08/2~/2011
(CONTINUED)
No AlB found
(NOAFBj
8/2(/2011
A~B CULTURE
AP'B CULTURE COlolMENT
P!tNDING
~re
inoculata<l (or
l~ entered at
week LJ C\,lltVl"e will
result
Dnl~
'l'950Q
SMEAR
EX~INATION
APB CULTURE
APB CULTURE
CAAD~N
COMM~NT
C.! RCUS J BO
ne~ative
6 weeks. Eight
Pe rspottea
if positive .
BRONCHIAL WASHLNGS
08/20/11
DBNNr S SCHMITT
No AF8 found
8/24/2011
(NOAFBJ
PE'.!'1D!NG
(!'Ollr lIl'li ts of lOediu!\\ eire inoculated for
Acid East
APAGE
CONTINUED
Advanced Diagnostic Labor&tories
NATIONAL JEWXSH HEALTH
1400 0ack~on Street
DENVER, CO
CAP #21789-01
l,:wllfr.:~
" . ,iJ,8'i
80206
ellA #00D0644307
LAB 106 (6,08)
1:47PM
T9500
AOx
TEST RES ULTS
NO, 6225
P. 6/6
(CONTINUED)
bci11i (AFB) ara read at 1,3. and 6
Default c~lt~re result is
" PENDING" ano should be interpreted
as NEGATIVE unti l a positive reault
is entered or the final neg~tive
result is entered at 6 wee~s . Eight
l\feek W culture will be reported
only if posit ~ ve\
we~s.
CARDEN C1RCUS,EO
3 PAGE
END OP REPORT
Advancea Diagnostic
La~oratories
:. ,;\!Wl!:~
. ' '~i!S'(
ellA lIoeD06443<l1
LAB 106 (8.0B)
~An .IWlS1/0flIV{~
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800 849 C869
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8/24/2011
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OB/19/2011 09;00
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08/19/11
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DEC-2e-2911 14r29
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IOtCF'IA NIAGARA
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TEST RSSUL.TS
08/19/2011 OBIOI)
PJ'B
00J5/0a5
o NatIonal JewIsh
Q Health'
':'9491
C1rcua Inc
(CONTINUED)
BOO 849 C86,
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812412011
CUlIIURZ tnX1AT:VE POR
10/06/2011
lofYCOIlACfl~IA
until a
po.~t!ve
r ult
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apgc:IiI!DI' SO[ffiC!
~CTI1AL- COLLEC'I' JAT:i
p~rIEt/'l'" S PRYBIC!AN
TRONK '/lASH
PKYBICtAH'S PHONE
H7 95'1 9Sn
800 949 0869
No ~~B ~our.d
PHYSICIAN) 8 F~ l'ID
Sl'IEAR !X.lY.INATlOr.r
C8/20/11
DENNIS aCHxITT
(~APB)
8/'34/2011
HAGel
CO~TINUED
JEWISH aEhLTH
Client
SeTv~ce~1
Le;endi H Hlgh
800~S50-6~2?
@. Critical
r.." Low
OEC-22-2el1 14:2B
IOI CFIA
NH~GARR
H21)
~tH
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f2.
20 J I
J1hll Gl7
an
I: 32P~l
T~492
v Health'
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TEST RESULTS
1YGI\\,NJubl.Ol'B
~1l1'\l.O.b 1D:h' 1Iho 1&
C~RDE~
C
I
(CDWINUSD)
"'i!
cut,rUllB NEGATIVE
10/06/2011
FO~
MYCOBACTERIA
I
I
bac!l~i
~d
....eGk.a.
FINAl REPOf( .
OEC 2220Hp~12\14
CARDEN
~~D
C!RC~S,SE~~
ci'Jl *211BH1
OF
R~?ORT
OEC-22-c8!1 14,21
91~1
!'HU 141
WICFIA NIAGAAA
~t
eLlA t/OBDDW3U1
!.AS 1;)6 (04.1')
I,:oll NO.
~H7'
~007
2/14/2012 8 : 24 AM
Pa.gs
of 7
. . . . . . .."\15 . . .".......
FINAL REPORT
:;r , . . ,
Ownor
AC0&89Ion Number;
11-063398
i98120144ge
OreanA County MO
Submitter 269lle
MO
Anlma( Location
(01
Collected By:
12/21/2011
02/13/2012
01. MIOhell1 Stal'(ord
Purpoa9:
TB10-4
PO BoXG
Fell Gre1/9, MO G664B
FAX #: 417-7697803
Phone #; 4177697900
Animal ID: 80 Case#: TB12-02037 Semple: 12/13/2011 Spealmen Typo: Trunk Washings Speclu
: Elflphanl, Aslan
No Isolal/on MS.de
Anlm.I 10: 80 Case #: T81202038 SamplG: 12114/2011 Specimen Typo: Trunk Washings Speoles
; Elephant, Asian
No Isolation Made
No Isolation Made
AnlmallD: Bimbo C(l6& #: TB1202040 Sample! 12/13/2011 8pocllTJon Typo: Trunk Washings
Species: E.lephant, Asian
No Isolation Made
AnlmsllD: Bimbo CaS9 II: TB1202041 Sample: 12(14/2011 SpecImen Type: Trunk Washings
Speoles: Elephant, Asian
No Isolation Made
Poge 1 of 4
Data Generelsd:
2/13/2012
211Q/2012 8:24 AM
Accession:
1H03398
Owner:
Page
George Cardell Circuli
4 of ?
Rafarrlll Number.
An Imal to: Bimbo Case #: TS 1202042 Sam pte: 12/15/2011 Specimen Type: Trunk Washings
Sp4!c19s~ Elephant, Asian
No Isolation Made
Anlm81lD: Janny Case #: T812-02043 Sample: 1211312011 Specllnen Wile: Trunk Washings
SpecleB: Elephant, Asian
No Isolation Made
Animal 10: Jenny Caso #: T81202044 Sample: 12/14/2011 Speelmon TYpo: Trunk Washings
Species: Elephant, Asian
Mycobacterial Culture Result
AnImal 10: Jenny Case #: lB12-Q2045 Sample: 12/1612011 Specimen Typo: Trunk Washings
Speolos; Elephant, Asian
Mycobacterial Culture Result
No lsolallon Made
Animal ID: Tory Case #: TB1202046 Sample: 12/13/2011 Specimen Type: Trunk Washings
Speoles: Elephant, Asian
Myoobacterlal Culture Result
No IsolaUon Made
AnlmallD: Tory CaS9 #: T91202047 Samplo: 12/1412011 Specimen Type: Trunk Washings
SFWoles: Elephant, Asian
Mycobacterial Culture ResuU
No Isolation Made
Animal ID: Tory C~M #: TB12-02048 Saml'le: 12115/2011 Specimen Type: Trunk Washings
Speolas: Elephant, Asian
Mycobaclerlal Culture Result
No 180lallon Made
Animal ID: Liz Case tI: T81202049 Sample: 12/13(2011 Speclman lYpa: Trunk WashIngs Species
: ElepMnt, AsIan
Mycobacterial Culture Result
No Isolation Made
Animal 10: Liz CalHI #: TB12.02050 Sample: 12114/2011 Speolmorl TYpe; Trunk Washings Speoles
: Elepha.nt, Asian
No Isolallon Made
AnlmallD: LIz Cau #: T812.02051 Sample: 12/1512011 Specimen Type: Trunk Washings Species
: Elephant, Asia/)
Mycobacterial Culture Result
No Isolation Made
Page 20(4
Date Gentllllted:
2113120 I2
2/14/2012 8:24 AM
Accession:
11-o83S98
Owner:
P~ge
5 Of 7
RelElrrnl Number:
AnlmsllD: Janice Case lJi T812-02052 Sample: 12/1312011 Specimen Type: Trunk Washings
Speoles: Elephant, Asian
No Isolallon Made
AnlmallD: Janice CSge #: T812-02053 Samplo: 12/14/2011 Speclrr19n Type: Trunk Washings
Spades! Elephant, Asian
Mycobaoterlal Culture Result
No Isotallon Made
AnlmallD: janice Cass #: TB1202054 Sample: 12/15/2011 Specimen TYpe: Trunk Washings
Species: Elephant, Asian
No
I~olatlon
Made
.----------------------~------------~
AnlmsllD: Cindy Case #: T81202055 Sample: 12(13/20'11
Specimen Type: Trunk Washings
Speclu$: Elephant, Asian
No Isolallon Made
AnImal 10: Cindy CaBe #: TB1202056 Sample: 12/14/2011 Specimen Type: Trunk Washings
S eoles: Elephant, Asian
Mycobacterial Culture Reeull
No Isolation Made
Anlma.IID: Cindy CSBe #: TB12 02057 Sample: 12/15/2011 Specimen Type: Trunk Washings
S oolos: Elepharn, Asian
No Isolallon Made
Animal 10: Vicky Case #: T8120205$ Sample: 12113/2011 Specimen TYP9: Trunk Washings
Speoles: Elephant, Asian
No Isolation Made
AnImal )0: Vicky Case #: T812-02059 Sample: 12/14/2011 Specimen Typ.a: Trunk Washings
SpGclos: Elephant, Asian
No Isolelion Made
Anlm<lllD: Vicky Caile #: TB1202000 Sample: 12.11512011 Specimen Type: Trunk Washings
No Isolatlon Made
AnlmallD: Belly Case #: TB12-02061 S~mple: 12/13)2011 Specimen Type: Trunk Washings
Speoles: Elephant, AsIan
No Isolation Made
Page 3 or 4
Date Generewd:
2/1312012
2/1~/2012
Accession:
11.063398
8:24 AM
Owner:
8 of 7
Page
Refs/1lI1 Number:
Animal 10: Betty Cllse #: TB1202062 Sample: 12/14/2011 Specimen IYP9: Trunk Washings
Spooles: Elephant, Asian
Mycobacterial Culture Rasul(
No Isoletion Made
Animal 10: Betty Ca8~ #: TB12-020e3 Sample: 12/15/2011 Specimen Type: Trunk Washings
SpeclQs: Elephant, Asian
No Isolal/on Made
AnlmallD: Judy CS80 #: TB1202064 Sample: 12/1312011 Specimen Type: Tfunk Washings
SpClcl9s: Elephant, Asian
Mycobacterial Culture Result
No Isolallon Made
~----------------------~---------~---------~--------------------.-------,
Animal 10: Judy C8S0 #: T81202055 Sample: 12/14/2011 SpBclm&n Type: Trunk Washings
Specills: Elephs1\t, Asian
No Isoletlon Made
Animal 10: Judy Case #: T812-02066 Sample: 12/15/2011 Specimen Type: Trunk Washings
Speoles: Elephant, Asian
No Isolallon Made
page 4 014
Date
Gonereted:
211312012
'
--V3 __
FINAL REPORT
. . '1~~~
1'..!'.. .......,....:...""
owner
1201B552
Accession Number:
George Carden
Iowa City, IA
NFC Control Number:
Datu Collected:
Dab! ReceJved:
Date ComplEllDd:
Collected By:
00511112012
07/09/2012
Dr. Allnda Buckingham
Purpose:
TS 10--4
Animal Location
Johnson County lA
SubmlUer- 27437
19612036;379
Specimen Type:
Trunk
No Isolation Made
AnimallD: 80 AVID# 036584056 Case #: TB1205791 Sampl&: 6/8/2012 Specimen Type: Trunk
Washings SpecIes: Elephant, Asian
No Isolation Made
Animal 10: 80 AVID# 036584056 Case #: T812 05792 Sample: 6/9/2012 Specimen Type: Trunk
Washings SpecIes: Elephant, Asian
. No Isolation Made
To W(pedlte the Pl'ocessing of youl' lostlng. pleaae use youI' Submltlcl' IIlouu(ei./ althe top IGIl oryow' rtlPOli un IIOY ,
future submIssions, and please remain consistent wllh your submission Infol'l11ollon (I.e, addre.~s, phone, fax, cmBI!, ele,),
Page 1 of 1
Date Generated;
71912012
.7/31/2012 9113 AM
Page
3 of 4
/"
't'
.....
~-~
_,..... ,,;,,
FINAL REPORT
PO 8o~ B44
Arne" Iowa 60010
Phone: 615-337-7614
Fait: 616-337.7938
FeDERAL RELAY SERVICE (VoJoe/TTYJASCIiISpanlah) 1-800.677.8339
The USDA IUh aqua( opponunHy provider andt)mp/oyer.
Owner
12-021322
AC088elon Number.
.SpringReld, MO
19812039342
Anlrnalloe.allon
Greene county MO
Oat. Colleoted:
Date Completed;
Col/ectad By:
Purpolle:
Submltt.r 26999
CJe/O 1120 12 .
07131/2012
Dr. Michael D. Stafford
Date Received:
T810-4
This la 8
b/lfablll O.lIe,
AnlmallD: Betty Catile #: TB1200108 8amplo; 5/23/2012 Spoclman Type: Trunk Washings
Specle8: Elephant
Mycobacterial Culture Result
No Isolallon Made
AnlmallD: Betty Case II: TB1200109 Sample; 5124/2012 Speclmliln Typo: Trunk Washings
Specla.: Elephant
No Isolation Made
nlmallD: Betty Csse #: T81200110 Sample: 5/25/2012 Specimen Typa: Trunk Washings
Speclea: Elephant
,haullS authorized
by:
No Isolation Made
~19 Uj tlQLR~
(TIl" new ~eD1lon "'" bl Up:!Olo!d p&rtodlOlify with l/pt for 9ubmftl&re.)
To oxp~/te the prOl"leMing DC yO\lr l681ing. pJeaso UJ& your
Submltl&r 1Ilocatod at
th6
Oil
any
fu~ 8ubmlsslollJ. IIlId plGUQ romaln coo.tlalonl with your subm/41iDD Informatioa (i,e. Iddr()4B. pIlona, fax. Dmall, etc.).
Page I 011
Date (3anarated:
7/31/2012
FINAL REPORT
A~cesllon Number:
12-033097
Springneld, MO
Animal Location
Greene Counly MO
Submitter' 26999
19812052182
08/28/2012
10124/2012
Dr. Michael D. Slof(ord
TB104
PO 60x 9
Thle 19 a billable
0&1$.
AnlmallD: 80 Case #: TB12-07936 Sample: 8/2212012 Spaclm9n Type: Trunk Washings Species:
Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
No Isolation Made
Animal ID: 80 Case II: T812-07938 Sample: 8/2412012 Specimen Type: Trunk Washings Speoles:
Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
AnlmallD: Betty Case #: TB12-07939 Sample: 8/22/2012 Specimen Type: Trunk Washings
Sp6clea: Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
AnlmallD: Betty Cas9 #: TB12-07940 Sample: 8/2312012 Specimen Type: Trunk Washings
No Isolation Made
Page 1 of 2
Dele Generated:
10124/2012
Accession:
12033097
Owner:
Relerrill
Number:
Animal 10: Betty Csse #: rB12-07941 Sample: 8/24/2012 Specimen Type: Trunk Washings
Species: Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
Animal 10: Bimbo Cue #: TB12-07942 Sample: 8/22/2012 Speolmen Type: Trunk Washings
Species: Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
Animal fD: Bimbo Case #: TB1207943 Sample: 8/23/2012 Specimen Type: Trunk Washings
Species: Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
Animal 10: Bimbo Case f#: T912-07944 Sample: 8/24/2012 Specimen Type: Trunk Washings
Species: Elephant, Asian
No Isolallol1 Made
.ubmlll.~ . )
To expedite the procc~lng of you I' testing, plc/lSe use your Suoml(ler ~ located at lhe top left of your repon on any
fuMe !ubmlsslons, Bnd pleDSC romain consls(onl with your submission informlllion (I.e. 6dclr~s, phol\e, fIIx, emoll, etc.).
Page 2 of 2
Date Generated:
10J2412012
\Ij)
\'.' !<.I IllMr
,
Labot'~tor1cs
. '..'{ ',
""ri
Ownll'l
Accoss/on NUIYlller.
12060113
Gl)orge Car<J~1l
Sprlngneld, MO
Anlmol Loootion
Greene COl.lllly MO
Sullm/ttG r 26999
19813013906
1212812012
03/05/2013
D,. Michael D. Sta/(ord
TB-104
CQ9\),
Animal 10 : Betty Ca&e #: TB1210503 Sample: 12/18/2012 Speclmon Typo: Trunk washings
Spao/es: Elephant, As/an
Mycobacterial Culture Result
No Isolation Made
Anlma/lD: Betty Case II: TB12-1050<1 Sample: 12/19/2012 Specimen Type: Trunk Washings
SpecIes: Elepi1an~ Asian
Mycobacterial Culture Resliit
No Isola(lon Made
Animal 10: Betty Csse #: T81210505 Sample: 12120/2012 Specimen Type: Trunk Washings
Speoles: Elephant, Asian
MycobacterIal Culture Result
No Isolation Made
fAnlmallO: 80 Case #: T812-10506 Sample: 12/18/2012 Specimen TYPQ: Trunk Wa!?hings Speclss
~ Elephant, Asian
._ _ ._________
No Isolallon Made
G'm~i-jD:-BO CElB;;;;:
: Elephant, Asian
No Isolation Made
Pag81 of 4
DctG GenerE1ted:
3/5/2013
Accession:
12.050113
Owner:
Gsorge Cardon
Relerl'lll Number:
Animal 10: Bo Case #: TB12-10508 Sample! 12/20/2012 Specimen Type: Trunk Washings Speoles
: Elephant, Asian
No 19o1ation Made
AnlmallD: Cindy Case #: TB12-10609 Sample: 12/18/2012 Specimen Typo: Trunk Washings
Speoles: Elephant. Asian
Mycobacterial Culture Result
No Isolation Made
AnlmallD: Cindy CeIIe~: TB12-10510 Sample: 12119/2012 Speclmlln Type: Trunk Washings
Species: Elephant. Asian
Mycobacterial Culture Result
No Isolation Made
AhlrnallD: Cindy Case #: TB12-105,1 Semple: 12120/2012 Specimen Type: Trunk Washlnge
Species: Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
Anlma/ID: Janice Caso #: TB12-10S12 Sample: 12/18/2012 Specimen Type: Trunk Washings
Species: Elephant, Asian
Mycobacterial Culture Result
Animal 10: Janice Case 'II:
Spe~le8;
No Isolallon Made
TB12~10513 Sample:
Elephant, Asian
No Isolation Made
Animal 10: Janice Cseo #: TB12-10514 Sample: 12/20/2012 Speolman Type: Trunk Washings
Speoles: Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
AnlmallD: Jenny Ca~e #: TB12-10515 Sample: 12118/2012 Speolmen Type: Trunk Washings
Species; Elephant, Asian
No Isolation Made
Animal 10: Jenny eaSD #: TB12-10518 Sample: 12/19/2012 SpeCimen Type: Trunk Washings
SpecIes:
Elaphan~
As/an
No Isolation Made
AnlmallD: Jenny Case tI: TB12-10517 Sample: 12/20/2012 Specimen Type: Trunk Washings
SpeclN: Elephant, Asian
Mycobacterial Culture Result
No IsolaOon Made
Page 2 cf4
Dale Generated:
3/612013
AW~S8lon :
i2'()50i13
Owner.
George carden
Referral Number.
AnlmallD: Judy Case tI: TB1210518 Sample: 1211812012 Specimen T~pe: Trunk Washings
Speole9: Erephant, AsIan
Conlaminated
Acid
Isolate
Mycobacterium trlvlale
The Isola Ie was sequenced allha1E1S rRNA and the rpoB regIons 01 the genome. Results were blasted agalns\
RIDOM and GenBank. The Isolate ma!ohed Mycobacterium !r!viale al 99% sequence Blmllarlty based 011 rpoB
sequenolng. Mycobacterium trlvlale belongs to Ihe M, terrae compl6X.
nlmal ID: JUdy Case #: T812-10520 Sample: 1212012012 Specfmen Type: Trunk Washings
Spocles: Elephant, Asian
No Isolatlon Made
Animal ID: Liz Case #: T81210521 Sample: 12/18/2012 Specimen Type: Trunk Washings Speoles
: Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
AnlmallD: Llz Cau8 #: TB1210623 Sample: 12120/2012 SpeCimen Type: Trunk Washings Species
: Elephant, Asian
No Isolation Made
Animal ID; Tory Case #: T81210524 Sample: 12/1812012 Specimen Type: Trunk Washings
SpecIes: Eleph8n~ Asian
No Isolation Made
AnImal ID: Tory Case #: T81210525 Sample: 1211912012 SpecImen Type: Trunk Washings
Specl86: Elephant, Asian
Mycobacterial Culture Result
AnlmallP: Tory Case
No laolallon Made
No Isolation Made
Page 3 ot 4
Date Generated:
31512013
AccessIon :
12-050113
Ownec
George Carden
Ratelral Number:
AnlmallD: Vicky Case #: TB12-10527 Sample: 12/1812012 Specimen Type: Trunk Washings
Species ; Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
AnlmallD: Vicky Case #: T612-10528 Sample: 12119/2012 Specimen Type: Trunk Washings
Species! Elephant, Asian
Mycobaclarlal Culture Result
No Iso\atlon Made
Anima! lD : Vicky Case #: T812-10529 Sample: 12120/2012 Specimen Type: Trunk WashIngs
Species : Elephant, Asian
No Isolation Made
.ubmi~e~)
To expedite the proCC3.'!lng of your sample(s), please Include Ihe word "sa m p!es" In tile addre~ on lite slllp
/0
labot.
Page" of4
Dale Generaled:
3/612013
_V;
FINAL REPORT
Owner
Accession Number:
Submitter' 28999
Collected By:
Purpose:
13023392
19813040149
0710212013
09103/2013
01'. Michael D, Stafford
TB-10--4
PO Box 9
Animal 10: 80 Case #: TB13-05046 Sample: 6/25/2013 Specimen Type: Trunk Washings Specles~
Elephant, Asian
Mycobacterial Culture Result
Isolate
AnlmallD: Bo Case #: T813-05047 Sample: 6/26/2013 Specimen Type : Trunk Washings Species:
Elephant. Asian
Mycobacterial Culture Result
No Isolation Made
An Imal 10: Bo Case #: TB13-05048 Sam pie: 6/27/2013 Specimen Type: Trunk Washings Species:
Elephant, Asian
No Isolation Made
AnlmallD: Betty Cllse #: TB13-05049 Sample: 6/25/2013 Specimen Type: Trunk Washings
Species: Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
Pags 1 of 2
Date Generated :
91312013
Accession:
13-023392
Owner:
Referral Number:
AnlmallD: Betty Case #: T813-05050 Sample: 6/26/2013 Specimen Type: Trunk Washings
Species: Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
Animal 10: Betty Csse #: T813-05051 Sample: 6/27/2013 Specimen Type: Trunk Washings
Species; Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
AnimallD: Vicky Case #: T813-05052 Sample: 6/26/2013 Specimen Type: Trunk Washings
Species: Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
AnlmallD: Vicky Case #: TB13-05053 Sample: 6/26/2013 Specimen Type: Trunk Washings
Species: Elephant, Asian
'
Mycobacterial Culture Result
No Isolation Made
Animal 10 : Vicky Case #: T813-05054 Sample: 6/27/2013 Specimen Type: Trunk Washings
Species: Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
Animal 10: Janice Case #: T813-05055 Sample: 6/25/2013 Specimen Type: Trunk Washings
Species: Elephant, Asian
No Isolation Made
Anima 110: Janice Case #: T913-05058 Sample: 6/26/2013 Specimen Type : Trunk Washings
Species : Elephant, Asian
No Isolation Made
AnlmallD: Janice Case #: T813-05057 Sample: 6/27/2013 Specimen Type: Trunk Washings
Results authorized by :
No Isolation Made
Sec~on
Halp Us Ha!pJ:.QJl
(1013 new .. ellen will be updsted ps(IOdlc.lly wlth UP$lor aubm!t!$I'(._)
Pleas6 remember to submit payment or your User Fee account number at the time of Bubmlsslon. If you
would like our billing department to contact you for a credit card number, please provide a valid contact
number,
Page 2 of 2
Dala Generated :
9/312013
f'C J
, "
/.
i (I
', ".
t ~ ,
'J
I' -,-.
~1("1
HN/Il RI'F'Un I
1:1,1
l-P,t..)IJ..jj'"
3:)..1Y
",,0)
()IMllf;'(
for u(ilcial
I):.;" lH Ily
13044382
(.;e()i'q( ~ (;11((\<-1'1
~~I'(il)yll"JII'I ;
MU
191j14(J10~39
Animal Locatloll
Gre[:ll\c Coul)ty MO
12/1 B12013
02/1 '112014
Data Compleled:
Cofleotacl By:
Dr, Michael
Purpose:
TB1D-4
Submlttfll' 26999
"J.'l'"'n.<f
Thle Is
D, StaNord
hlllllbftl case.
Washings Species:
E:lephal1'l, Asian
_ _ '_ ,_ _ , _ ,.. A". _
_
, .~-
, .~
'.,
_~~
..... _. , ___ .
_, _ , _ _ _
~ _.
_ _ _ _ _ _.. _ _ _ _
.. _ .
~_,_._J
No Isol~lI!)n Made
Wa~hin9::>
___ ., __ .... '-
_ ~
_.~.
Mycoh~lcler'iElJ
..;._~._,
Culture Result
. __ . _ _ _ , _
...'-"' _ _ _ _ _ _ _ _ _ _ ., _ _ _ . _ . _ _ _ _ _ _ _ _ _ _ .' __ _
No Iso/Eltion Made
:J
__._-_._._--_.--' __
_._--_.__.-_.
.._---,---_ .....__ .
..- ..
..
AnJmalID: B'3(ly, 036570847 CaBfI #: TB13 ..08189 SamplG: 121'111'1:1 Specimen Type: Trunk
!IN'1shil1as
-, -- ~ ~
, - - -
Sr~H:los :
- - _._- , , _ ... -
- - -- -
I--
,. -- .~ - ~'~. -'-
-,
No Isol<ltion Made
-_'__ .. - .... _."--- ,_._-_._---_ .. ....------,, .- ._.. _...
Af)rm~IID : 801ly, 038;-i19847 C,\!?6 #; TEl'l:.1-08HJO Sample: 121'12/13 Spe<:lmen Type: Trunk
lNasrlings SpecJl.s : Eleph"ml, Asian
-_._---...'---,,- .,-,
.... . ,-----, . . - "-, ... -_.- - .. ,.- --, .......
~-
E=.II-lph<mlc I\siClIl
-'"' '
---_...
_....
_ ._-"
_._-
_-_
-- . ~-.~-
-,
---
_._-_._--_
__ _._]
.._...
No Isolation Made
O~te G Clrum.rtecl:
2}'1'11201A
..
..--.". -
Accession:
1J04131l2
Owner:
George Carden
Referral Number:
Animal 10: Betty, 036579847 Case #: TB13Q8191 Sample: 12/13/13 Spec Imen Type: Trunk
Washings Species: Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
AnlmallD: Cindy, 036549535 Case #: T81308192 Sample: 12111/13 Specimen Type: Trunk
Washings Species: Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
AnlmallD: Cindy, 038549535 Case #: TB1~08193 Sample: 12/12/13 Speelman Type: Trunk
Washings Species: Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
Animal 10: Cindy, 036649535 Case #: TB13-0S194 Sample: 12/13/13 Specimen Type: Trunk
Washings Species: Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
AnlmallD: Janice, 4999761240 Case fl.: TB13-08195 Sample: 12/1'1/13 Specimen Type: Trunk
Washings Species: Elephant, Asian
No Isolation Made
AnImal ID: Janice, 4999761240 Case #: TB13-08196 Sample: 12/12/13 Specimen Type: Trunk
Washings Species: Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
An/mal 10: Janice, 4999761240 Case "Ii: T613.08197 Sample: 12/13/13 SpecImen Type: Trunk
Washings SpecIes: Elephant. Asian
Mycobacterial CultUre Result
No Isolation Made
Animal [D: Jenny, 036323638 Csse #: TB13-08198 Sample: 12/11/13 Specimen Type: Trunk
Washings Species: Elephant, Asian
Mycobaoterlal Culture Result
No Isolation Made
Animal 10: Jenny, 036323638 Case #: T813-08199 Sample: 12/12/13 Specimen Type: Trunk
Washings Species: Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
An/maIID: Jenny, 038323638 Csse #: TB13-08200 Sample: 12/13/13 Specimen Type: Trunk
Washings Species: Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
Page 2 of 4
Dale Generated:
2111/2014
Accession:
13044382
Owner:
Referral Number:
George Carden
Anima I 10: LIz Case #: T81308201 Sample : 12111/13 Specimen Type: Trunk Washings Species:
Elephant. As ian
Mycobacterial Culture Result
No Isolation Made
Anima! 10: Liz Case #: TB13-08202 Sample: 12/12/13 Specimen Type: Trunk Washings Species:
Elephant, Asian
Mycobacterial CultUre Result
No Isolation Made
A nlm al 10; Liz Case #: T813-08203 Sample: 12/13/13 Specimen Type: Trunk Washings Species:
Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
Animal ID: Tory, 4A18064623 Ca88 #: TB13-Q8204 Sample: 12/11/13 Specimen Type: Trunk
Washings Species: Elephant, Asian
Mycobacterial Culture Re!;lull
No Isolation Made
Animal 10: Tory, 4A18064623 Case #: TB13-08205 Sample: 12/12/13 Specimen Type: Trunk
Washings Species: Elephanl, Asian
Mycobacterial Culture Result
No Isolation Made
Animal 10: Tory, 4A18064623 Case #: T8130S206 Sample: 12/13/13 Specimen Type: Trunk
Washings Species: Elephant, Asian
MycobacterIal Culture Resu lt
No Isolation Made
Animal 10: Vicky , 036560042 Case #; TB13-08207 Sample: 12/11/13 Specimen Type: Trunk
Washings Species: Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
AnlmallD: Vicky, 038580042 Case #: TB1308208 Sample: 12112113 SpecImen Type: Hunk
WashIngs Species: Elephant, AsIan
Mycobacterial Culture Result
No Isolation Made
Animal 10: VIcky, 036560042 Case#: T813-08209 Sample: 12/13/13 Sp6clmen Type: Trunk
Washings SpecIes: Elephant, Asian
Mycobacterial Culture Result
No Isolallon Made
Page 3 o( 4
Dale Generated:
2/1112014
Accasslon:
13-044382
Owner:
George Carden
Referral Number.
Please remember to BI.lbmlt payment or your User Fee account number at the lime of submission. If you
would like our blUing department to contact you ror a credit card number, please provide 8 valid contact
number.
Page 4 of"
DElte Generated:
211 112014
_. __,_.~ug
27
2014
HP LASERJET FAX
lO:OBAH
p, 1
FINAl. REPORT
B~
844
Filii: 513-3J7.T938
, FEDERAL RELAY SERVICE (VolaefTTY/A8CIIIBp.~I.n) ~ . 60o-&17.8339
The U80A ~ .lin equal Opportl.llllly pro~i<Ior 'lind amploy.r.
OW"lr
Cerdsri Clroua
Springl1C1ld, MO
Ma.nlon Humber:
NFO Conlnll Number:
Dati Col18ct8d:
1ge14034311
Greene county MO
D"t, A'.'v.d:
o.t. Ollmpl.l.d:
08(2D1'2.014
0811912014
liIubmll~r
Anlm.' L.ocatlO"
14..()20098
26999
Purpo.. ;
No IsolallOI1 Made
No laolatlon Made
Ima' ID! 90 Cae. ,: TB14.()2463 Sample: 6/18/2014 Speo lme" Type: Trunk Wuhlnga Speclea:
Ele han'. Asian
No Isolation Made
nlmallD: Be/tty CU. #: TB14-D2464 Sample: 6/16/2014 Sp.clmen Type: Trunk WQshlng8
SpecIes: Elephant. Asian
No laolst!on Made
," nl rna I to: BlJtffy C . . . II: T814..Q2486 Samp'e: 6/17/2014 _peclme" Type: Trunk Washings
No laolatlon Made
Page 1 or 2
Dst" 06neral.,o:
(JOB NO.
e.t1912l1t4
6517)
IalOOl
Au~
27 2014 IOIOBAN
HP LRSERJET
FA~
Referrul Numbor:
nlmallO: 8eltty Can fr T814-0246El Simpla: 6/16/2014 Speolmen Type: TrunK Washings
Elephant, Asian
epec~.:
No laolatlon Made
Anlmell to: Vicky Cue #: TB1402467 Sample: 6/16/2014 Specimen Type: Trvnk Washings
Specl : Elephant, Asian
Mycobacterial Culture Reaull
No Isolatlon Made
"'maIIO: Vloky en.~: TB1402468 8emple: 6/1712014 9peolm.n Type: Trunk Washings
Specl. . : Elephant, Asian
MycobactarJal Culture RElsuH
No Isclatlon Made
nfmall0: Vicky Cu. AI: TB14024e9 Semple: 6/18/2014 SpecImen Type: Trunk Weshlnge
Speol : ElephBnt, Asian
Mycobaoterial Culture Result
No laolstlon Made
crnl I'>CIIir Hotlon ~Ibe updated p91iodb111y wflh tIPoI for wbmlt""")
Quality lIamples yiBld thllllTlost aoourate I'e!lulle. PleaSE: call If you htlve qU9!1tfo ns .
page 2 0(2
08/27/2014
Det, Gilnenallld:
no
10113
8/19/2014
""002
USDA
FINAL REPORf
Owner
AC08Sslon Number.
15000343
Goorge Carden
Springneld, MO
Animal Looatlon
Greene Counly MO
Submitter. 26999
Fair Grove Vel Service
PO Box 9
Fair Grovel, MO 65648
Dale Colleoted:
Date Roo elved:
Date Comploted:
Colleoted By:
Purpoae:
01/06120,5
03/02/2016
Or, Michael D. Siafford
T8-10-4
FAX'll: 4177!!9-7803
Phone II; 417 -769-7803
This I, not a billable case,
AnlmallD: Bo Case #: TB15-00032 Sample: 1/02/2015 Specimen Type: Trunk Washings Species:
Elephanl. Asian
No Isolallon Made
Animal ID: 80 Case #: T815-00033 Sample: 1/03/2015 Spaclmen Type: Trunk Washings Species:
Elephant. Asian
No Isolation Made
AnlmallD: Bo Case #: TB15-00034 Sample: 1/0512015 Specimen Type: Trunk Washings Species:
Elephsnl, Asian
No Isolation Made
Animal ID: Betty elise #: T815-00035 Sam pie: 1/0212016 Specimen Type: Trunk Washings
Species: Elephant, Asian
No Isolation Made
AnImal ID: Betty Case #: TB 15-00038 Sample: 1/03/2015 Specimen Type: Trunk Washings
Spocles: Elephant, Asian
No Isolation Mede
Page 1 of 4
Dale Generated:
31212015
Accession:
16.000343
Owner:
Goerge Carden
Referral Number:
Animal 10: Betty Case #: T815-00037 Sample: 110512015 Specimen Type: Trunk Washings
Species: Elephant, Asian
Mycobacterial Culture Resull
No Isolation Made
Animal 10: Janice Case #: T815-00038 Sample: 1102/2015 Specimen Type: Trunk Washings
Species: Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
AnlmallD: Janice Case #: T81500039 Sample: 1/03/2015 Specimen Type: Trunk Washings
Species: Elephant, Asian
Mycobacterial Culture ResuU
No Isolation Made
Animal ID: Janice Case #: TB15-00040 Sample: 1/0412015 Specimen Type: Trunk Washings
Species: Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
AnlmallD: Vicky Case #: T81500041 Sample! 1/02/2015 Specimen Type: Trunk Washings
Species: Elephanl, Asian
Mycobacteria! Cutture Result
No Isolation Made
Animal 10: Vicky ClIse #: T815-00042 Sample: 1/0312015 Specimen Type: Trunk Washings
Species: Elephanl, Asian
Mycobacterial Culture Result
No Isolallon Made
AnlmallD: Vicky Case #: T81500043 Sample: 1/04{2015 Specimen Type: Trunk Washings
SpecIes: Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
AnlmallD; Cindy Caso #: T81500044 Sam pie: 1/02/2015 SpecImen Type: Trunk Washings
SpecIes: Elephant. Asian
Mycobacterial Culture Resull
No Isolation Made
llimallD: Cindy Case #: TB15-00045 Sample: 1/03/2015 Specimen Type: Trunk Washings
Species: Elephant, Asian
Mycobacterial CultUre Result
No Isolation Made
AnlmallD: Cindy Case #: TB16-00048 Sample: 1/05/2015 Specimen Type: Trunk Washings
Species: Elephant. Asian
Mycobacterial Cullure Result
No Isolation Made
Page 2 of 4
Date Generated:
31212016
Accession:
16000343
Owner:
Goerge Carden
Referlai Number:
AnlmallD: Tory Case #: T815-00047 Sample: 1/02/2015 Specimen TyPB : Trunk Washings Species
: Elephant, Asian
Mycobacterial Cullure Result
No Isolation Made
AnlmallD: Tory Case fl.: TB15-00048 Sample: 1/03/2015 Specimen Type: Trunk Washings Species
: Elephant, Asian
Mycobacterial Culture Result
No Isolallon Made
Animal 10: Tory Case #: TB15-00049 Sample: 1/05/2015 Specimen Type: Trunk Washings Species
: Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
AnlmallD: Jenny Case fl.: TB1S-00050 Sample: 1/0212015 Specimen Type: Trunk Wa9hlngs
Species: Elephant, Asian
Mycobacterial Culture Result
No Isolalion Made
Animal fO; Jenny CU4;7 #: TB15-00061 Sample: 1/03/2015 Specimen Type: Trunk Washings
Species: Elephant, Asian
No lsolallon Made
Animal 10: Jenny Case #: TB 15"00052 Samplo: 1/04/2015 Specimen Type: Trunk Washings
Species: Elephant, Asian
Mycobacterlel Culture Resull
No IsolaHon Made
AnlmallD: Liz Csse #: TB15-00053 Sample; 1/0212015 Specimen Type: Trunk Washings Species:
Elephant, Asian
No Isolation Melde
AnlmallD: liz Cane #: T815-00054 Sample: 1103/2015 Specimen Type: Trunk Washings Species:
Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
AnlmallD: Llz Case #: TB15-00055 Sample: 1/05/2015 Specimen TYPB: Trunk Washings SpecIB&:
Elephant, Asian
Mycobacterial Culture Result
No lsolatlon Made
Page30r4
Dale Gef)eral6d:
31212015
Access ion:
1B-000343
Owner:
Goerge Carden
RelerrBI Nt/mbet:
Quality samples yield the most accurate results. Please ca ll If you have questions.
Page 4 ot 4
Dale Generated:
31212015
USDA
FINAL REPORT
POBox 844
Ames, loWl! 60010
Phone: 615-3377514 Fox: 615337793B
FEDERAL RELAY SERV1CE (VolcaffTY/ASCIIISparJlah) 1-800...977-8339
Tho USDA Is an equal opportunity provider and employer.
Owner
Animal Location
Submltt&r - 26999
Fair Grove Vet Service
PO Box 9
Fair Grove, MO 65848
FAX # : 417-759-7803
Phone #: 417-759-7803
Accession Number.
NFC Control Number:
Date Collected:
Date Received :
Date Completed:
Collacted By;
Purpose:
15-019796
19815034330
OB119/2015
08/1812015
Thhs Iii
billable cue.
No Iso!ahon Made
nlmallD: Betty Case #: TB15-02604 Sample: 6/1412015 Specimen Type: Trunk Washings
Species: Elephant, Asian
Mycobacterial Culture Result
An ImalIC: Belty Case #: T815-02605 Sample : 6/15/2015 Specimen Type: Trunk Washings
Species: Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
AnlmallD : Cindy Case #: TB15-02606 Sample: 6/1212015 Specimen Type: Trunk Washings
Species: Elephan~ Asian
Mycobacterial Culture Result
No Isolation Made
AnimallD: Cindy Case #: T815-02607 Sample: 6/14/2015 Specimen Type: Trunk Washings
SpeCies: Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
Page 1 of2
Date Generaled:
8118/20i5
Accession:
15-019796
Owner:
Referral Number:
AnlmallD: Cindy Case #: TB15-02608 Sample: 6/15/2015 Specimen Type: Trunk Washings
Species: Elephant, Asian
Mycobacterial Culture Result
AnlmallD: Bo Case #:
Elephant, Asian
T815~02609
No Isolation Made
Sample: 6/12/2015 Specimen Type: Trunk Washings Species:
No Isolation Made
AnlmallD: Bo Case #: TB15-02610 Sample: 6/14/2015 Specimen Type: Trunk Washings Species:
Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
Animal ID: Bo Case #: TB15-02611 Sample: 6/15/2015 Specimen Type: Trunk Washings Species:
Elephant, Asian
Mycobacterial Culture Result
Help Us
No Isolation Made
Help You
(This new section will be updated periodically with (Ips fOT subml~8(&.)
Quality samples yield the mosl accurate results. Please call If you nave questlons,
Page 2 of 2
Date Generated'
8/18/2015
Good afternoon,
Thank you for sending the additional documentation regarding Betty and Bo. The DCHHS has reviewed and notified us
that their position has not changed and they will not be able to be exhibited at Unlversoulln Dallas , Communication Is
attached.
That being said we can continue to process your event permit without Betty and 80 or If you would like to use
alternatlve animals. Please advise as soon as possible so we can coordinate efforts . As a reminder, If you plan to use
additional elephants/animals at the event the following documents will be required :
'.
---------------
EXHIBIT 3
Subject: Unlver$oul Circus (Letter to City of Dallas RE Elephants reactive to TB) - 2015 Dallas Animal Permits
Importance: High
Ms, Ausherman :
Last month while the circus was in Norfolk, Virginia, UnlverSoul Circus provided trunk wash test results to the Norfolk
Deputy City Attorney responsible for animal welfare, to counter claims made In June that Bo and Betty have been
positive for TB. Also Included in the documentation was a letter from Dr, Stafford, the elepha nts' veterinarian, and a
detailed explanation of the types of TB tests and USDA guidelines for TB for elephants. All of the documentation and full
statement are attached to this e-mail. In particular, I want to share with you the following excerpts from the June 15,
2015 correspondence from our Norfolk counsel to the Norfolk Deputy City Attorney, copied below:
I am sending YOll this in response to your expression of concern abollt whether two of the elephants currently traveling
with the UnlverSoul Circus (now In Norfolk) have tuberculosis. The two elephants at Issue are Betty and Bo. They
belong to larry Carden, who Is the trainer traveling with them. Your concern Is premised on your having been sent DPP
testing results for Betty and Bo (and other elephants owned by Mr. Carden) from December of last year. Those test
results show samples taken on December 9,2014 using a DPP test resulting In a "Reactive Ab Positive" finding In a report
dated December 23,2014. Since we spoke about this Friday afternoon, I have spoken with Mr, Carden, Dr. Michael
Stafford (the primary veterinarian for Bo and Betty, and the person who submItted the DPP samples at Issue In
December), and several officials at UnlverSoul Circus about the elephants' health, the December TB screen, and the
safety of their traveling and being exhibited here In Norfolk.
To be clear, I am assured by Dr. Stafford that neither Betty nor Bo have TB. He could not have been any more emphatic
about that Moreover, Dr. Stafford provided me the attached statement, dated Friday, June 12, that attests to Betty and
Bo meeting or exceeding the relevant USDA guidelines for the control of TS in elephants. A copy of his statement Is
attached ,
I am also attaching the USDA's own testing results of the more accurate trunk wash tests for TB done on both elephants,
both before and after the December DPP screening at Issue. The first of those was based on samples taken on June 16,
17, and 18, 2014. The more recent was based on samples taken on January 2,3, and 5, 2015. The reported finding of
"No Isolation Made" means that no positive TB cultures could be Isolated from the samples tested. It Is the way that a
negative trunk wash TB test result is reported by the USDA.
The complete correspondence from our lawyer to the City of Norfolk Is attached . It comprehensively (overs the TB Issue
as it relates to Betty and Bo, and should be sufficient evidence to allow Bo and Betty to perform In Dallas.
You can reach me at 404-588-1235 ext 8730 or Ruthline Kelly at 404-588-1235 ext 8734 with any questions. Also, Dr.
Michael Stafford has made himself available to discuss any concerns or questions as well.
Ben Johnson
Soul Circus, 'nCo
NEW ADDRESS
230 Peachtree Street, NW
Importance: High
Hello,
I am reaching out to let you know we have denied the permission to exhibit Betty and Bo, your two elephants at the
UniverSoul event; as well as any other animal that has tested r2actlve to tuberculosis . Please see the attached
communication from Dallas County Health and Human Services.
Please let me know as 500n as possible If you plan to use any additional elephants at the event. The following
documents will be required asap If are planning replacements:
Please contact me directly should you have any questions. Thank youl
Zachary Thompson, MA
Director
Sincerely.
jf1:m7~~
Director, Dallas County Health and Human Services
C: Dr. Christopher Perkins, Dallas County Medical DirectorlHealth Authority
_____ __
~
..
, _
_______ "
______
__
__
__ A
[ am reaching out to let you know we have denied the permission to exhibit Betty and Bo, your two elephants at the
UnlverSoul event; as welJ as any other animal that has tested reactive to tuberculosis. Please see the attached
communication from Dallas County Health and Human Services.
please let me know as soon as possible If you plan to I,Jse any'addltlonal elephants at the event. The follOWing
documents wHI be required asap If are planning replacements:
1
EXHIBIT 4
Please
DCHHS
Safe
Zachary Thompson, MA
Director
July 27,2015
Joey Zapata
Assistant City Manager, City of Dallas
Via email: joey.zapata@dallascityhall.com
Ron King
Exocutive Director of Convcmlion and Event Services, City of Dallas
Via email: ron .king@dallascityhalJ.com
Kris Sweckard
Director of Code Compliance, City ofDailas
Via emall; kris .sweckard@dllllilScilYhal1.com
Dear Mr. Zapata, Mr. King, and Mr. Sweckard,
Dallas County recently received infOlmation indicating tbat two elephants scheduled to appear with Soul
Clrcus, Inc. (also known as UnlverSoul Circus) at the Soulhw6Sl Center Mall in Dallas next month have tested
reactive for tubercUlosis. Dallas County Immediately reached out to the Toxas Oeparunenl of State Health
Services and the United States Department of Agliculture to discuss the potential risks associated witb
bringing these elephants mto (he area.
Based on our communications with these agencies, Dallas County Medical Dil'ectorlHeelth Authority Dr.
Christopher Perkins who, by agreement, serves as the Health Authority for the City of Dallas, Dallas County
Director of Tuberculosis Control Dr. Garry Woo, Imd I respectfully request and recommend that tbe City of
Dallas deny Soul Circus, Inc. permission to exhibit these elephants, Betty and Bo, liS well as any other elephant
that has rested reactive fur tuberculosis. Given the potential transmission of tuberculosis from elephants that
CIIl'ly the disease to arena and circus workers, members of the public, and other animals, we believe that these
elephants should not be exhibited.
Should Soul CIrcus, Inc. seek to bring other elephants to the event scheduled in August, please request the
following infolmatlon and forward it to my office:
copies of the elephants' medical records;
photographs of the elephants;
infDlln8tion regarding any Identffying marldngs of the elephants; and
the name of the cDmpany's onsite veterinillian.
Please contact me If you have any questions.
Sincerely,
g~m~o~
Subject: UnlverSoul Circus (Letter to City of Dallas RE Elephants reactive to TBl - 201S Dallas Animal
Permits
Ms . Ausherman:
Last month while the circus was In Norfolk, Virginia, Un IverSoul CIrcus provided trunk wash test results
to the Norfolk Deputy City Attorney responsible for animal welfare, to counter claims made In June that
Bo and Betty have been positive for TB. Also Included In the documentation was a letter from Dr.
Stafford, the elephants' veterinarian, and a detailed explanation of the types ofTS tests and USDA
guidelines for TB for elephants. All of the documentation and full statement are attached to this email. In particular, I want to share with you the folloWing excerpts from the June 15, 2015
correspondence from our Norfolk counsel to the Norfolk Deputy City Attorney, copied below:
I am sending you this In response to your expression of concern about whether two of the elephants
eu rrently travel ing with the UnlverSoul Circus (now In Norfolk) have tuberculosis. The two elephants at
Issue are Betty and Bo. They belong to Larry Carden, who Is the trainer traveling with them . Your
concern Is premised on your having been sent DPP testing results for Betty and Bo (and other elephants
owned by Mr. Carden) from December of last year. Those test results show samples taken on December
9, 2014 using a opP test resulting In a "Reactive Ab Positive" findl ng In a report dated December 23,
2014. Since we spoke about this Friday afternoon, I have spoken with Mr. Carden, Dr. Michael Stafford
EXHIBITS
(the primary veteri narlan for Bo and Betty, and the person who submitted the DPP samples at issue In
December), and several officials at UnlverSoul Circus about the elephants' healthJ the December TB
screen, and the safety of their traveling and being exhibited here In Norfolk.
To be clear, I am assured by Dr. Stafford that neither Betty nor Bo have TB. He could not have been any
more emphatic about that. Moreover, Dr, Stafford provided me the attached statement, dated FrldaYJ
June 12J that attests to Betty and Bo meeting or exceeding the relevant USDA guidelines fDr the control
of T8 In elephants. A copy of his statement Is attached.
am also attaching the USDA's own testing results of the more accurate trunk wash tests for TB done on
both elephants, both before a nd after the December DPP screening at issue. The first of those was
based on samples taken on June 16, 17, and 18,2014. The more recent was based on samples taken on
January 2, 3, and 5, 2015 , The reported finding of "No Isolation Made" means that no positive TB
cultures could be isolated from the samples tested. It Is the way that a negative trunk wash TB test
I
the
City
You can reach me at 404-588-1235 ext 8730 or Ruthline I(el'y at 404-588-1235 ext 8734 with any
questions. Also, Dr. Michael Stafford has made himself available to discuss any concerns or questions as
well.
Ben Johnson
Soul Circus, Inc.
NEW ADDRESS
230 Peachtree Street, NW
Suite 2000 - 20 th Floor
Atlanta, GA 30303
404- 787 -1821 mobile
bj ohnson@universoulclrcus.com
Fro m:
Ausherma n,
Ivery, Rhonda
Importance:
High
Hello.
[ am reaching out to let you know we have denied the permission to exhibit Betty and Bo, your two
elephants at the UniverSoul event; as well as any other animal that has tested reactive to
tuberculosis. please see the attached communication from Dallas County Health and HUman Services.
Please let me know as soon as possible if you plan to use any additional elepha nts at the event. The
following documents will be required asap If are planning replacements:
Please contClct me directly should you have any questions. Thank youl
DCHHS
g,~m~C~<U
being held up in litigation. To further c:ompllca te things, the 2008 gUidelines call for the use of the STAT PAK, but-as
noted above-It Is no longer available for use. So the current state of affairs Is that the 2008 guidelines are the effective
ones, but they have to be appl\ed without their STAT PAK requirement. The draft 2010 gUidelines call for use of the opp
test, but the DPP test has not yet been approved by the USDA. Mr. Carden stili uses It anyway, because some states
require It. The USDA gUidelines call for an annual trunk wash test, but Mr. Carden has that test done every six months as
a precaution that goes over and above what the law requires. I am told that the next trunk wash for the elephants with
the UnlverSoul Circus now In Norfolk is due to be conducted this weel<.
Under the USDA guidelines} an elephant should not travel whlle.l.t has TB or If It becomes "untested". An elephant
becomes I/untested" If It ~oes for more than 12 months without a negative trunk wash test being performed. Hence,
given all of that, there Is no reason why Betty and Bo cannot travel toes'e days.
Finally, you said you were concerned about the potential for an elephant with TB transmitting the disease to humans.
a m Informed by Dr. Stafford that no one really knows whether humans can contract TB from an ele'phant. To date, there
has been one suspicious case In the U.S., but no one has yet been able to prove that that person contracted T6 from an
elephant or, fra nkly, whether anyone could. Evidently, It Is much more likely that the transmission could go the other
way: that humans may be able to transmit T6 to elephants.
As I had indicated to you Friday, my understanding Is that this Is not the first time that PeTA has sought to use the TB
scare as a way to frustrate the Inclusion of elephants In traveling circuses.
Ho pefully. the Information provided above and In the documents attached wUI serve to dispel the notion that either
Betty or Bo have TB or pose any kind of public health rlsl<. Nevertheless, If you need any additional Information or
documentation In that regard, my client and I will be more than happy to provide It to you right away. - Chris
Christopher Abel
WILLCOX & SAVAGE, P.C,
440 Monticello Avenue, Suite 2200
Norfolk, VJrglnla 23510
Direct Dial: (757) 628-5547
Direct Fax: ("757) 333-3647
Cell: (757) 870-5344
. This message Is confidential, Intended only for the named reclplent(s) and may contain Information that Is privileged,
attorney work product or otherwise protected by applicable law. If you have received this message In error, please notify
the sender at 757-628-5500 and delete this message.
USDA
Bo~
FINAL REPORT
844
II
Aceeulon Number:
14-020098
Carden CirCUli
Springfield. MO
Animal Location
Greene Counly MO
Submitter - 26999
Fair Grove Vel Service
PO Box 9
Fair GrovB, MO 65648
FAX #: 417-769-7803
Phone #: 417-769-7603
19814034317
06125/2014
Collected By:
08119/2014
Dr. Michael D. stafford
Purpose:
T8-10-4
This 18
II
billable case.
Animal 10: Bo Case #: TB14-024B1 Sample: 6/16/2014 Specimen Type: Trunk Washings Species:
Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
AnimallD: 80 Caso #: TB14-02462 Sample: 6/17/2014 SpecImen Type: Trunk Washings Species:
Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
Animal 10: 80 Case #: T814-02463 Sample: 6/18/2014 Specimen Type: Trunk Washings Species:
Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
Animal 10: Bettty Case #: TB14-02464 Sample: 6/16/2014 Specimen Type: Trunk Washings
Species: Elephant. AsIan
Mycobacterial Culture Result
No Isolation Made
AnlmallD: Bettty Case #! TB14-02465 Sample: 6/17/2014 Specimen Type: Trunk Washings
SpecIes: Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
Page 1 of 2
Dilta Generated:
8/1912014
Accession:
14-020098
Owner.
Cardan Circus
R~ferr81 Number.
AnlmallD: Betlty Case #: TB14-02486 Sample: 6/1812014 Specimen Type: Trunk Washings
Species: Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
AnlmallD: Vicky Case #: TB14~02467 Sample: 6/16/2014 Specimen Type: Trunk Washings
Species: Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
AnlmallD: Vicky Case #: TB14-02468 Sample: 6/17/2014 Sp8clm~n Type: Trunk Washings
Species: Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
AnlmallD: Vicky Case #: TB14-02469 Sample: 6J18/2014 Specimen Type: Trunk Washings
SpecIes: Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
QualHy samples yield the most accurate results. Please cal! If you have questions .
Page 2 of 2
Data Generated :
8/1912014
USDA
FINAL REPORT
PO Box 844
AmM, Iowa 50010
Phone: 615-337-7614 Fax: 515-337-7938
FEDERAL RElAY SERVICE (VolcafITY/ASCIl/Spanlsh) 1-80Q...S77-fl339
The USDA Is an equal opportunity provider and employer.
PO Box 9
Fair Grove, MO 65648
FAX #: 417-769-7803
Phone #: 417-759-7803
15-000343
Accesslorl Number:
Date Collected:
01/0612015
0310212016
Dr. Michael D. Stafford
TB-10-4
Date Received:
Date Completed:
ColIQcted By:
Purpoae:
RllfOrTilllRetalrl Tag Number:
CIISB.
Animal 10: Bo Case #: TB15-00032 Sample: 1/02/2015 Specimen Type: Trunk Washings Species:
Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
AnlmallD: Bo Case #: T815-00033 Sample: 1/03/2015 Specimen Type! Trunk Washings Species:
Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
Animal 10: 80 CasEI #: TB15-00034 Sample: 1/05/2015 Specimen Type: Trunk Washings Species:
Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
AnlmallD: Betty Case #: TB15-00035 Sample: 1/02/2015 Specimen Type: Trunk Washings
Species: Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
AnlmallD: Betty Case #; T81500036 Sample: 1103/2015 Specimen Type: Trunk Washings
Species: Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
Page 1 of 4
Date Generated:
3/2/2010
Accession:
16000343
Owner.
Goerge Carden
Referral Number.
Animal ID: Betty Csse #: TB15-00037 Sample: 1/05/2015 Specimen Type: Trunk Washings
Species: Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
Anlm81lD : Janice Case #: T815-00038 Sample: 1/02/2015 Specimen Type : Trunk Washings
Species: Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
Anlm81lD: Janice Case #: T815-00039 Sample: 1/03/2015 Specimen Type : Trunk Washings
Species: Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
Animal ID : Janice Case #: TB15-00040 Sample: 1/04/2015 Specimen Type : Trunk Washings
Species: Elephant. Asian
Mycobacterial Culture Result
No Isolation Made
AntmsllD: Vicky e8Se #: T815-00041 Sample : 1/0212015 Specimen Type: Trunk Washings
Speclee: Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
AnlmallD: Vicky Case #: T815-00042 Sample: 1/03/2015 Specimen Type: Trunk Washings
Species: Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
No Isolation Made
AnlmallD: Cindy Case #; T615-00044 Sample: 1/02/2015 Specimen Type: Trunk Washings
Species: Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
Animal 10: Cindy Case #: T815-00045 Sample: 1/03/2015 Specimen Type: Trunk Washings
Species: Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
Animal 10: Cindy Case #: TB15-00046 Sample: 1/05/2015 Specimen Type: Trunk Washings
Species: Elephant, Asian
Mycobaclerial Cu lture Result
No Isolation Made
Page 2 of 4
Date Generated :
3/212015
Accession:
16-000343
Owner:
Goerge Carden
Rarerral Number.
AnlmallD: Tory Case #: T815-00047 Sample: 1/02/2015 Specimen Type: Trunk Washings Species
: Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
Animal ID: Tory Case #: T815-00048 Sample: 1/03/2015 Specimen Type: Trunk Washings Species
: Elephant, Asian
Mycobacterial Culture Result
No Isolatlon Made
AnlmallD: Tory Case #: T815-00049 SlImple: 1/05/2015 Specimen Type: Trunk Washings Species
: Elephant, Asian
Mycobacterial Culture Result
No Isolalion Made
AnlmallD: Jsnny Case #: TB15-00050 Sample: 1/02/2015 Specimen Type: Trunk Washings
Species: Elephanl, Asian
Mycobacterial Culture Result
No Isolation Made
AnlmallD: Jenny Case #: T815-00051 Sample: 1/03/2015 Specimen Type: Trunk Washings
Species: Elephant, Asian
Mycobacterial Culture Result
No Isolatlon Made
AnlmallD: Jenny Case #: TB15-00052 Sample: 1/04/2015 Specimen Type: Trunk Washings
Species: Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
AnimallD: Liz Case #: TB15-00053 Sample: 1/02/2015 Specimen Type: Trunk Washings Species:
Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
AntmallD: Liz Case #: T815-00054 Sample: 1/03/2015 Specimen Type: Trunk Washings Species:
Elephant, Asian
Mycobacterial Culture Result
No Isolation Made
Animal ID: Liz Case #: TB15-00055 Sample: 1/05/2015 SpecImen Type: Trunk Washings Species:
Elephant, AsIan
Mycobacterial Culture Result
No Isolation Made
PBge 3 of 4
Date Generated:
31212015
Acceseion:
16-1] 00343
Owner:
Goerge Carden
Referral Number:
Hilip
U8
Help You
Quality samples yield the most accurate results. Please eaUlt you have questions.
Page 4 of 4
Dale Generaied:
312(2015
DCHHS
~~ lives.
Dallas County Health and Human Services
Safe fa.Milies.,
Zachary Thompson, MA
Director
Kris Sweckard
Director of Code Compliance, City of Dallas
Via email: kris.sweckard@dallascitvhall.com
Re: Soul Circus Inc. elephants
Dear Mr. Zapata, Mr. King, and Mr. Sweckard,
Thank you for sending us the additional information from Ben Johnson with Soul Circus, Inc.
regarding the company's desire to exhibit two elephants that have tested reactive for tuberculosis
at an event in Dallas next month.
Dallas County Medical DirectorlHealth Authority Dr. Christopher Perkins who, by agreement,
serves as the Health Authority for the City of Dallas, Dallas County Director of Tuberculosis
Control Dr. Garry Woo, and I have reviewed the recently submitted information and our position
stands . While we recognize that the individuals charged with caring for these elephants have
differing viewpoints, our investigation shows that the threat of potential transmission of
tuberculosis from elephants that carry the disease to arena and circus workers, members of the
public, and other animals remains a serious concern. We believe no elephant that has tested
reactive for tuberculosis should be exhibited.
Sincerely,
j~m&S~
Director, Dallas County Health and Human Services
2377 North Stem mons Freeway, Suite 600, Dallas, Texas 75207-2710
Office 214.819.2100 Fax 214.819 .6022
EXHIBIT 6
TE~
S U ITED
FAX: 770.410.32.81
~PHAR~A.GA300D9
770A10.1555
WWW.BRISKINLAW.COM
August 17,2015
VIA E-MAlL TO
SARAH.AUSHERMAN@DALLASCTTVHALL.COM AND
REGULAR
U.S. MAIL
In short, the analysis provided by DCHHS is flawed and fails to recognize or give
proper scientific weight to the indisputable evidence that neither of the elephants, Bo nor
Betty, has TB. Further, the analysis fails to recognize that, as of the date of tlris letter,
there has never been a single reported case of transmission of TB from an eJephant 0 a
member of the general public arising from any public exhibition of elephants.
From the letter provided to us from the DCHHS, it appears that the sole basis for
the recommendation of the denial of the permit for exhibition of the elephants Bo and
9USINESS .
TECHNOLOGY.
COMMUNITY.
E)(HIBIT7
&
SANFORD, LLC
AT LAVJ
Betty is an unfounded fear that any elephant that has ever "tested reactive of
tuberculosis" poses a threat to the public. Such is simply not the case .
Enclosed with this letter, please find a copy of the supporting materials for the use
of the Cherubio Diagnostic Systems, Inc. DPP Vet TB Assay for Elephants. As the
literature clearly states, in the first paragraph, "The test is used as an aid in the diagnosis
of active tuberculosis (TB) in conjunction with other diagnostic tests." [11]
(Emphasis added.) The literature further states "A reactive result is suggesti~/e of active
TB" (2J (Emphasis added.) Of most importance is the following statement, found in the
third full paragraph : "The only USDA-recommended definitive diagnostic test to
detect TB in live elcpbantg is mycobacterial culture of trunk wash samples." [3] As
such, the DPP reactive immunochromatographic rapid test is merely a cbeap, easy, quick
screening tool, for which a follow up trunk wash is the only definitive test The DPP test
is subject to false positives, and the USDA guidelines, which are binding authority,
approve of trunk wash testing as the only defmitive test for active TB in elephants. As
such, any decision based solely on an old DPP false positive that was subsequently
proved to be a false posi tive by subsequent negative trunk washing tests ignores the best
scientific and medical evidence without any basis, and as such, is umeasonabJe, arbitrary,
and capricious. This is all the more so, when one considers that Betty and Bo meet all of
the requirements of both the USDA and the State of Texas [or exhibition in your state.
Soul Circus' elephant vendor, in an abundance of caution and in appreciation of the need
to fonow the best scientific evidence available, [ollowed the 20]0 Guidelines for the care,
testing, diagnosis, and treatment of Betty and Bo, the elephants in consideration for this
permit. This testing and care was provided and supervised by Michael D . Stafford,
D .V .M., who will testify t:hat he and tile owner of the elephants have fully complied with
the most restrictive protocols related to these animals, the 2010 USDA Guidelines.
I t is important to note that, under the 2010 Guidelines, elephants that test reacti ve
or positive to a DPP test, but also have a negative culture from a trunk wash and no
U $
I r..;
s _
TECH NOL OG Y .
A T L. A W
known exposure to TB fall under Group 3C . The 20 I 0 Guidelines prescribe how often
elephants that fall under Group 3C should be tested . The 20 I 0 Guidelines specifically
state that elephants should be "[mJonitor[ed] by culture (triple sample method) every 3
months for one year, then every 6 months fOT life. " The 2010 Guidelines further state
that elephants should undergo "repeat MAPIAlDPP testing every 3 ' months for the first
year, then every 6 months for an additional 2 years if elephant remains . .. reactive. If
all cultures remain negative after 3 years annual serological testing may resume as
described in these guidelines." The 2010 Guidelines then go further to state that " the
elephant may be able to travel and have public contact after 6 months of successful
documented USDA approved treatment."
The very first time that Bo tested reactive for TB was a Stat-Pak test in 2009, and
the very first time Betty tested reacti ve for TB was a State-Pak test in 2011. Immediately
after receiving these results) the staff and veterinarians in charge of the care of Betty and
Bo perfonned trunk wash tests every 3 months for one year (as prescribed by the 2008
and 2010 Guidelines), which all came back negative, and have been performing trunk
wash tests every 6 months thereafter (as prescribed by the 2010 Guidelines), which have
all come back negative for TB. Since all cultures remained negative for 3 years, annual
serological testing resumed. Currently, there are no restrictions on travel and exhibition
for Bo and Betty under the 20 10 Guidelines. As such, the Circus believes that it has fully
complied with the most restrictive suggested guidelines for treatment and testing.
Accordingly, the Circus suggests tbat the refusal of the City of Dallas Office of Special
Events to issue the requested permit is without scientific or legal basis,
As the materials provided to you previously have demonstrated, the trunk wash
tests performed on January 2,3, and 5, 2015, are all negative for Bo and for Betty. The
reported findings demonstrate that no positive TB culture could be isolated from the
series of samples . Under the '2010 Guidelines, these consistently negative tests mean
both elephants are under no restriction or special protocol of any kind and-in
accordance with even the most conservative interpretation of the applicable rules-may
be treated as if they never had a positive test. Consistent with those testing results, it
should be noted that the elephants' veterinarian has emphatically stated that neither one
has TB . The Circus previously has provided all of that evidence to your office.
While we certainly understand-and share-the City's concern over the
transmission of tubercuJosis, we also believe that the exclusion of Bo and Betty is
unreasonable, arbitrary, and capricious. It is clear from the correspondence from the
DCHHS that the 2008 Guidelines and more restrictive 2010 Guidelines were not
rev iewed or consulted in making tltis decision. As such, we believe that the decision is
based on less than a full Wlderstanding of the veterinary medical evidence, and, in fact,
ignores the clear and indisputable scientific evidence. Accordingly, we respectfully
hereby request that your office reverse its previous decision and grant the Circus the
requested perrni1s for the exhibition of both elephants. We will be happy to engage in
SU t ; I N E S $
TECH NOLOGY.
City of Dal1
Pnge 4 of4
any dialogue or to provide any other infonnation, including the names of recognized
experts in the field, should you so desire.
Given the reliance upon the exhibitor's compliance with the even more stringent
requirements of the 2010 Guidelines, the Circus is uncertain as to the legal or scientific
basis for the Department's denial. Please advise us of the specific statutory, regulatory,
or scientific basis for the deniaJ so that the Circus can evaluate an appropriate course of
action. The July 301h letter from DCHHS that was sent to the Circus merely states that
"our investigation shows that the threat of potential transmission of tuberculosis from
elephants that carry the disease to arena and circus workers, members of the public, and
other animals remains a serious concern." The letter from DCHHS does not state any
basis for their concern.
If you have any questions or would like to discuss this matter in more detail, feel
free to contact me at my office.
Sincerely yours,
BRISKIN, CROSS
~~<jl?Z
Alan M. Briskin
Attorney at Law
Enclosure
celene:
Mr. RanKing
Executive Director of Convention and Event Services, City of Dallas
Mr. Kris Sweckard
Director afCade Compliance, City of Dallas
TECHNOLOGY.
-C 0
!1 M U N I T Y
CATALOG II 65-9110-0
2 Tc.sl Kit
'HEMBIO
OIAGNOSTIC
SYSIEM5,
INC .
PRINCIl'LE OF TEST
TNTEND ED USE
The DP? VorTB Assay is a slJ1gJe use Immunooh romalographic rap id teat for Ihe del~tion of andbodios to
M)lCobacler (~m I~berclrlo:rls and Mycobacleril/n! Mvls In
serum, plasma or whole blood from Ati1c~ elephants
(LoxodontrJ afrloana) and Asian elephants (Elephas
max/mil!). The test is usod iIB an aid in the dlagnosb of active
lubercuJ05il1 (TB) In conjunction with other dl!gnostlc
methods.
nOll/'\laetive.
MATERIALS PROVIDED
Each kll contBins the follOWing lre~ :
IO -61 9~O
Product Insert
Rov 2
The Chembio DPP VelTB Assay Is b(\$ed on immunochl'Olnt\tographlc technology. The ~I emplo)13 two
reC<lmbinanl antigens, an M 11iberClJlwl~ and an M. bovls
antigen, which are separately Immobilized on tho membrane
solid phase. It also utilizes recombinant Protein AlG corUugBtlld
to colloidal gold pel110les for antibody detection. Ths DPP
VetTB AJse.y \lIIe.!I !erurn, plasma or whole blood. The ample
is applied to the SAMPLE+BUF'PER well with the buffer.
After the sample and buffer have mJgrated onto the rut strip
additional buffer ~ ~dded to the BUFFER well. The buffer
feolUtalll.'l tho lawai flow of the rulcnsod produc!:l IUld
promote.! the binding of antibodies to the antigens. If present,
the antibodies bind to !he gold conjugated antibody binding
prott:ln. In a I'CIIctive sample, tha dye conJugated-lmmu~e
complex migrates on tbe nitrocellulose mtmlbrnne find is
CJIpTUred by the antigens immobilized In the TEST (\ 2) area
producing 8 plnklpwple line. In Iho absence of detectable
IDltibody, no speclfio Immune complex would be formed on the
test line. Rnd, therefore, no pinklpurplo line would appear In the
TIlST (J 2) I\relI. Unbound cOrU\lgBted gold pllrtlcles continue
to ro!grate illong the rnem bra no I!lld produc6 a pink/purple Hne
In the CONTROL (e) area. This procedural control serves to
dcmonltr8te tluu rhe reagents havo been properly IIppUed end
have migrated through ll1e dovico,
hS. I of4
TEST PROCEDURE
2.
PRECAUTIONS
I.
2.
3.
4.
S.
6.
7.
8.
J,
1\110
We!12
SAMPLE +SUFFER
Weill
U~\ng a 10)1.1
4.
5,
6.
SPECIMEN COLLECTION
The DPP VetTB AS.!iay can be performed on whole blood,
serum or plasm8.
Whole Blood: Collect whole blood into tubes containing
10-6294-0 Rov 1
N<rVembe: 2012
7.
8.
Nonreactive Res\llt
Only n pink/purple CONTROL (C) line is visIble. The sample
oonllllns no deteclalJle antibody to both TB lind
mycobacteriosis antlgons. A nonreactive resull doe.! nol
preclude the possibility ofTB infection.
waslo conl~lner.
QUALITY CONTROL
A pinldpurple color~ IIno should always appear in
CONTROL (C) eruu if the test has been performod correctly
and the devico Is worldng proporly, It sarve.9 as I1n interNlI
test proccdul-al control.
Invalid Result
INTERPRETATION OF RESULTS
Reactive ResD It
l. Three pink/purple lines. one line In the CONTROL area.
one line 10 the TBST (l) area and one line in the TEST (2)
area indicates a reactlvo resu It. This SUggOBI3 that the
slImple is raACtive for TB .
2.
3.
4.
5.
3. A plnklpurple TEST (I) line and II plnk/pul'ple CONTROL
line 81'e vl~lb Ie, This suggests that th~ sampla is CtIIIotiyO ror
TB or mycobActeriosis.
TBST (ljUn.
6.
7.
8.
9,
speclll1on.
10. A nOIl-reaotlve result does not preclude the pos:libllity of
exposure to TB or Infection with TB. An antibody response
10 recent exposure may take several months to reaoh
detectablo levolg.
I!. In h~Bted elephants, Inle'lJl'et results with caution:
e, Treatment against T8 may reduce antibody responge.s.
thuB resulting ill non-reactive result:; in solllO c~ses.
Pes, J of4
I'ERFORMAi'lCE CHARACTERISTICS
732.
Greenwald, R. L)llI!Ihchenko, 0 ., fufundiari, J., Miller,
M., Mikota, S., Olsen, I.H., "Ball, R., Dumonc5Bux, G.,
Schmitt, D" Moller, T" Payeur, 1.B., Han'is, B" SofnHlko,
D., Wate~. W,R., Bnd L),a3hcbenko, K,P. (2009) Highly
accurate antibody assays fo!' easly Bnd rapid detection of
tubcl'Culosis in Aft'iean and ,A.,lan elephants. Clinical And
Vaccine immunology /6:605-612,
LYBShchenko K.P., Ore~nwBld R., Bsfilnd 1llTi J., Mlkola
S., Millor M.. Moller T., Vogc)nest L., Gtlrhe K.P ..
Rabbe-AustelmlUl S., Gal J., Dnd Walers W.R, (2012)
Field applJcBllon of serodlagnostlCB to idenllry elephl!1lts
who tubl:l'CLllosls prior to case confirmation by culture.
elin. Yacclne immltno/. [9(8):1269-75.
3.
4,
Elephllllt
species
Amcan
AfrlcBn
Mycooactel'ial
Asirul
Tobie 2.
Speclnci ~
Spe1::le5 Isolated
DPP VetTB
reactive
M tuberculMI9
616
M. boyls
1/1
33/3 3
M tuberculoJ/J
8rudl~
Elophant
ap5ci1;3
Atriono
79/79
A91an
68/68
Trunk wash
Mgative
79/79
cultur~
68168
REPRODUCIBILITY STUDIES
Reproduoibillty was evaluated at three independent
Illbol1ltories using two serials of DPP VeITB AasIl.Y. A
r8feren~ plDlel of 40 blindly-coded samples representing
negative, weakly reactive snd strongly reactIve Bera Wl."1'l!
lMled 3 tim68 on J different daYJ. The compliM re!ul\li from
J labol'elories demonstt'llted 98.6% acoUf8cy ,
REFERENCES
I.
2.
I D.4I294 0 R"" 2
NOV'embcr 20\2
Pag.~.fd
Cet#
Product
65-9[ [0-0
65-91 [1-0
65-9112-0
II
Elephant TB Guidelines 1
EXHIBIT 8
Elephant TB Guidelines 2
1. INTRODUCTION
Tuberculosis (TB) is caused by bacteria in the genus Mycobacterium. Over 100 species
comprise this genus. Mycobacteria infect a broad range of species including humans, nonhuman primates, carnivores; marine mammals, psittacine birds, reptiles, fish, artiociactylids,
pachyderms, and domestic and non-domestic ungulates. Species susceptibility to specific
mycobacteria varies (Montali 2001).
In mammals, the term ''tuberculosis'' is used to define disease caused by Mycobacterium
tuberculosis (M tb) complex organisms. The M tb complex includes M. tuberculosis, M. bovis,
M. africanum, M microti, M . canetti, M. caprae, and M. pinnipedii. A vaccine strain derived
from M. bovis (M bovis BCG) is sometimes included as a separate member of this complex.
The term "mycobacteriosis" refers to infection with any mycobacteria but is generally used to
defme disease caused by non-tuberculous mycobacteria (NTM). "Atypical mycobacteria" or
"mycobacteria other than TB" (MOTT) are other tenns used to describe this group. Most NTM
are saprophytes found in soil or water but they may occasionally cause disease in humans and
animals, including elephants .
2. DEFINITIONS
Ancillary diagnostic test: A subordinate OT auxiliary test to be used in support of a primary test
to diagnose disease.
Airborne transmission. Airborne transmission occurs by dissemination of either airborne
droplet nuclei or small particles in the respirable size range containing infectious agents that
remain infective over time and distance (e.g., spores of Aspergillus spp, Mycobacterium
tuberculosis bacilli). Microorganisms carried in this manner may be dispersed over long
distances by air currents and may be inhaled by susceptible individuals who have not had faceto-face contact with (or been in close proximity to) the infectious anima.l or person (Siegel 2007).
Elephant TB Guidelines 3
Attending veterinarian: a person who has graduated from a veterinary school accredited by the
American Veterinary Medical Association's Council on Education, or has a certificate issued by
the American Veterinary Medical Association's Council on Education Commission faT Foreign
Veterinary Graduates; has received training and/or experience in the care and management of the
species being attended; and who has direct or delegated authority for activities involving animals
at a facility subject to the jurisdiction of the Secretary (i.e. a USDA licensed facility).
Atypical mycobacteria: see non-tuberculous mycobacteria
Contact transmission:
Direct contact transmission may occur during activities such as touching or riding an
elephant, being touched by an elephant, examining, medicating, bathing, and handling
Indirect contact transmission involves contact with a contaminated intermediate object, such
as occurs during cleaning cages and equipment and handling soiled laundry. Injuries from
contaminated sharps, such as scalpel blades, needles, and necropsy knives, may result in
exposure to pathogens . (NASPHV 2006)
Culture positive for M.th complex: Isolation and identification of M. tuberculosis complex
organisms from any site using standard mycobacterial methods.
Culture positive (M.th complex) elephant: An elephant from which a M. tuberculosis complex
organism has been isolated from any body specimen. A culture positive elephant is considered
positive until it has met the treatment requirements as outlined in the CWTent Guidelines.
Dual Path Platform (DPP) VetTB Assay: A new generation screening kit for the rapid
detection of IgG antibodies to M tuberculosis or M. bovis in elephant serum, plasma, or whole
blood. The DPP has shown 100% correlation with MAPIA TM (Greenwald et a1. 2009).
ElephantTB STAT- PA.l( Assay: A qualitative screening kit for the detection of antibodies to
M tuberculosis and M bovis in elephant sera, plasma, or whole blood (Lyashchenko 2005, 2006,
Greenwald 2009).
ELISA: Enzyme-linked irnmunosorbent assay; a test used to detect and measure either antigen
or antibody.
Exposure: Risk of transfer of an infectious agent from 8 TB infected elephant(s) or
contaminated environment through contact (direct, indirect) or airborne modes of transmission.
Fondte: An inanimate object or material on which disease-producing agents may be conveyed.
Gamma-interferon test: A whole blood in vitro assay that can be used as an ancillary
diagnostic test for TB (not currently available for use in elephants).
Genotyping assay: A technique for the identification and analysis of polymorphism in certain
Elephant TB Guidelines 4
types of repeat units in DNA. Restriction fragment length polymorphism (RFLP) and variable
number tandem repeat (VNTR) are examples of genotyping techniques.
Herd: A group or groups of elephants, maintai.ned on common ground. Alternatively, two or
more groups of animals under common ownership or supervision that are geographically
separated, but that may have an interchange or movement of animals or personnel without regard
to health status.
Incidence: The rate at which a certain event occurs, for example, the number of new cases of a
specific disease occurring during a certain period.
Index animal: The animal in which a disease is first diagnosed.
Infected elepbant: an elephant from which Mycobacterium tuherculosis complex has been
identified through culture, PCR or other molecular techniques or that is reactive on the
ElephantTB STAT -P AK@ Assay and the MAPIA TM.
Intradermal tuberculin test (skin test): The injection of purified protein derivative (PPD)
tuberculin into the skin for the purpose of detecting exposure to tuberculosis. In cattle, the test
site is either the caudal fold (CFT) or cervical region (e.g. comparative cervical test, CCT) and
the test is read by observation and palpation at n hours (plus or minus 6 hours) following
injection. In hwnans, the test site is the foreann and the test is read at 48-72 hours . The
intradermal tuberculin test is not a reliable test in elephants (Mikota 2001, Lewerin 2005) .
Licensed veterinarian: a person who has graduated from an accredited school of veterinary
medicine and who has a valid license to practice veterinary medicine in the U.S.
MultiAotigen Print ImmunoAssay (MAPIA TM): A confIrmatory test to the ElephantTB
STAT -P AK Assay for detection of antibodies to M tuberculosis and M bovis in elephant sera
or plasma (Lyasbchenko 2000, 2006, Greenwald 2009).
Mycobacteria otber tban TB (MOTT): See non-tuberculous mycobacteria.
Mycobacteriosis: A disease caused by non-tuberculous mycobacteria (NTM).
Mycobacterium bovis (M. bovis); The primary causative agent of tuberculosis in cattle, bison,
and cervids; may also affect a variety of mammals including pigs, humans, primates, and nondomestic ungulates.
Elephant TB Guidelines 5
may also affect a variety of animals, including primates, pigs, cattle, dogs, parrots, elephants, and
rhinos.
Elephant TB Guidelines 6
3. ANNUAL TESTING
To adequately address the concerns ofTB in the general elephant population, all captive
elephants must be tested annually by culture and with the ElepbantTB STAT-P AK Assay (a
blood test). Samples for cultures and blood must be collected by or under the supervision of a
licensed veterinarian according to current USDA requirements. Blood collection for the
Guideline-required ElepbantTB STAT -P AK Assay must be witnessed by a federal or state
veterinarian and performed as licensed by the USDA Center for Veterinary Biologics. See
further information below under ElephantTB STAT -P AK Assay. It is required that elephants
with a reactive ElephantTB STAT-PAK Assay result be tested using the confirmatory
MultiAntigen Print ImmunoAssay (MAPIATM). See item 5 below.
Elephant TB Guidelines 7
Elephants should be tested within 30 days of the established annual test date. Blood for
ElephantTB STAT -P AK Assay and culture should be collected within a 2 week period. AU
elephants must be tested every calendar year. Note that the date the sample is collected is the
"test date," the date the sample is received at the laboratory is the "submission date," and the
date the laboratory reports the results is the "report date."
Record keeping of TB testing and treatment by the attending veterinarian is of upmost
importance. It is recommended that attending veterinarians maintain open communication with
the United States Department of Agriculture (USDA) and State Veterinarian, particularly
concerning elephants under treatment for TB or in cases of exposure to TB positive elephants . It
is recommended that at least a 1 ml aliquot of sera collected at the time ofTB testing be sent to
the elephant serum bank (See appendix 8).
C. Forcible exhalation - Mucous collected without instilling saline into the trunk is acceptable
if elephants are trained to forcibly exhale into a clean plastic collection bag and the volume
collected is at least 20 ml. This may allow sampling of secretions from other areas of the
respiratory tract and may be a preferable sample. Transfer the sample into sterile, leak proof,
plastic screw-top container.
Elephant TB Guidelines 8
Storage
Do not expose samples to sunlight or heat. Consult receiving laboratory to determine whether
samples should be refrigerated or frozen prior to shipment. For those laboratories that
recommend freezing (i.e. NVSL) freeze samples as soon as possible after collection and keep
frozen until shipment. Freeze at -20C (conventional freezer). As standard frost-free freezers
undergo cyclic freeze-thaws to limit frost, freezers that do not have this feature are preferred.
Freezing at -80 cC (ultra-low temperature freezer) is also acceptable. Frozen samples must be
shipped within 2 weeks of sample collection to the testing lab.
Elephant TB Guidelines 9
5. ELEPHANTTB
PROCEDURE
STAT~PAK~
Blood collection for the Guideline-required ElephantTB STAT -PAK Assay must be witnessed
by a federal or state veterinarian and performed as licensed. It is advisable to also bank a serum
sample. Blood from elephants with reactive ElephantTB STAT -P AKoo Assay results must be
submitted for MAPIA TM /DPP testing to:
Cherubio Diagnostic Systems, Inc.
3661 Horseblock Road
Medford, NY 11763
Tel: 631-924-1135
Fax: 631-924-6033
Email: customerservice@chembio.com
Contact Chembio for shipping instructions.
The USDA veterinarian is responsible for shipping the sample but the owner must pay for
shipping and must contact Chembio to arrange payment for the MAPIATM or DPP@ test.
Elephant TB Guidelines 10
STAT-PAK test should be repeated in 3 months time to confirm. If the ElephantTB STATP AK@ remains non-reactive, the elephant continues in Group 1.
known exposure
Monitor by culture (triple sample method) every 3 months for the ftrst year after
becoming ElephantTB STAT-PAK reactive, then every 6 months for the next 2
years. Repeat MAP LA TM /DPP@ every 6 months for the first year if elephant
Elephant TB Guidelines 11
Elephant TB Guidelines 12
Monitor by culture (triple sample method) every 3 months for one year postexposure, then every 6 months for life. Repeat MAPIA TM/DPP every 3 months for
the first year, then every 6 months for an additional 2 years if elephant remains
STAT-PAK reactive. If aU cultures remain negative after 3 years, annual
serological testing may resume as described in these Guidelines.
No travel or public contact until the first year of testing has been completed.
Treatment should be considered. If serological conversions are demonstrated to be
recent (within the past 12 months) then prophylactic treatment can be used. If
serological conversions are longer standing or unknown then full treatment may be
advisable. Individual cases should be evaluated in conjunction with USDA. If
treatment is perfonned, the elephant may be able to travel and have public contact
after 6 months of successful documented USDA approved treatment.
If the culture or MAPIA TM/DPP@ results change during any of the follow-up testing
the individual elephant will change group.
Culture positive elephants that have completed a course of anti-tuberculosis therapy
may remain ElephantTB STAT -PAK reactive and fall into this category. If
appropriate treatment has been documented and approved by USDA, these animals
will not have travel/public contact restrictions unless there is a change in their results
during follow-up testing. It has been shown that the MAPIA TM/DPP@ will decline
and may indicate a response to treatment so on-going annual monitoring with
MAPIA TM/DPP is required for life as changes in MAPIA TM may detect relapse.
Elephant TB Guidelines 13
on risk. If culture results during any of the follow-up testing become positive, the individual
elephant wj]l move to Category 4.
Consideration should be given to minimizing or eliminating contact with the public tha.t would
result in exposure by contact or aerosol transmission and to providing personal protective
equipment such as a NIOSH certified N95 respirator 1N95 face mask for staff when working in
close proximity to elephants that are under enhanced surveillance. Employees must be respirator
fit tested before they use the N95 respirator.
Based on a history of exposure to a culture positive animal, or other considerations, the attending
veterinarian may elect to administer prophylactic or full treatment after consultation with USDA.
Effective prophylactic therapy is defined as the administration of a specific number of doses of
two anti-TB drugs within a specified time. It must be demonstrated that adequate anti-TB drug
levels are achieved in the blood of the elephant under treatment. Acceptable anti-tuberculosis
drugs include isoniazid (INH), rifanlpin (RIP), pyrazinamide (PZA), ethambutol (ETH), or a
fluoroquinolone such as levofloxacin, moxofloxacin, ciprofloxacin, or enrofloxacin. Isoniazid is
recommended as one of the two drugs if a known exposure case isolate is INH sensitive. PZA
should not be given if M. bovis infection is suspected since this organism is inherently resistant
to PZA.
Prophylactic therapy is for 9 months can be administered using either of the following
schedules:
Prophylactic Treatment Schedule 1 (preferred):
Administer two anti-TB drugs daily for 9 months (270 total doses). The ftrst 60 doses should be
administered within a period of90 days (i.e. no more than 30 days of "refused medication"
should occur. It must be docwnented that the elephant received 270 total doses at a dosage level
sufficient to achieve adequate drug serum levels.
Prophylactic Treatment Schedule 2:
Administer the two anti-TB drugs daily for two months (as above, the first 60 doses should be
administered within a period of 90 days) . Adequate levels of both drugs must be demonstrated
in two serum samples collected approximately two weeks apart. Serum samples should be
collected as soon as the elephant is accepting medication reliably. If acceptable levels (see
below) are not achieved, the dosage should be adjusted and serum levels tested again (two
samples collected approximately two weeks apart) . It must be documented that the elephant
received the first 60 doses at a dosage level sufficient to achieve adequate drug serum. levels.
Once this has been demonstrated, administer the two drugs every other day but at twice the
previous dosage level for an additional 9 months (105 total doses of every other day dosing plus
the initial 60 doses for a total of 165 doses). It is not necessary to repeat serum drug levels when
changing to the every other day schedule.
Note: Pyridoxine 50 mg is administered to humans receiving INH for treatment of active or
latent tuberculosis to prevent the development of peripheral neuropathy. Although this side
effect has not been reported in elephants, it may be possible. At the discretion of the attending
veterinarian, Vitamin B6 (pyridoxine) can be given prophylactically at a dose of 0.8-1 mglkg
Elephant TB Guidelines 14
daily.
Concomitant use ofINH, rifampin, and PZA with other hepatotoxic drugs should be done with
caution.
Refer to TB Drugs section for starting dosages, routes of administration, side effects, blood
levels, and other information.
Monitoring of Prophylactically Treated Elephants
During the 9 months of treatment, elephants should be closely observed for changes in appetite,
behavior, and any other signs that may be attributable to adverse drug effects . Monthly blood
tests (CBC and serum chemistry profile) are recommended to monitor general bealth and
possible drug effects on the liver. Liver tests (AST, ALT, LDH, bile acids, and bilirubin) should
be included in the serum chemistry panel. Isoniazid may cause hepatitis and anemia. In
addition, leukopenia has occurred in at least one elephant apparently due to JNH toxicity).
GROUP 4: M. tuberculosis complex positive culture
Animals that have had Mycobacterium tuberculosis complex isolated from any sample (sputum,
stool, tissue, etc .) are considered culture positive for TB. A culture positive elephant is defmed
as an elephant from which Mycobacterium tuberculosis complex organism has been isolated
from any body site or specimen..
The ElephantTB STAT~PAK and MAPLA TM/DPP tests must be performed on blood from
culture positive elephants. Serum for MAPIA TM/DPP testing must be submitted regardless of
ElephantTB STAT -P AK resu Its .
Positive cultures must be submitted to NVSL for genotyping.
A culture positive elephant is considered positive until it has met the treatment
requirements as outlined below. These elephants must be separated from the public for the
duration of the treatment period. Separation from previously non-exposed elephants is also
recommended_until treatment is completed. Precautions to safeguard personnel health and safety
should be instituted immediately (see Employee Safety and Health section) . Elephants with
cultures that yield non-tuberculous strains of mycobacteria are not considered infected and are
not a risk to other animals or humans. Options for Category 4 elephants include:
Options:
A. Treatment: This is the preferred option for culture positive elephants whenever possible.
1. If the organism was isolated at a l.aboratory other than NVSL and they do not perform
mycobacterial species differentiation and DNA fingerprinting, the owner must request that the
laboratory submit the isolate to NVSL or other qualified laboratory for mycobacterial species
differentiation and DNA fingerprinting.
2. Antimicrobial sensitivity testing should be perfonned on all positive isolates. Sensitivities
should be requested for the following drugs: isoniazid, rifampin, pyrazinamide, ethambutol,
ciprofloxacin (or other fluoroquinolone), and amikacin. (Antimicrobial susceptibility testing for
Elephant TB Guidelines 15
Elephant TB Guidelines 16
Travel: Elephants in Group 4 should not travel or have public contact (direct or indirect) until
treatment is completed according ta the guidelines.
Group 5: Untested If an elephant cannot complete procedures as outlined far official annual
testing, it should not be permitted to have public contact that would result in exposure by contact
or aerosol transmission, or contact with other tested elephants (or their enclosures or equipment).
Untested elephants should not be moved from their home facilities. A tested elephant should not
move into a facility housing an untested elephant unless it can be demonstrated that there will be
no direct contact with the untested elephant or with its enclosure or equipment. If a tested
elephant(s) is in contact or housed with an untested elephant, the tested elephant carolOt travel
nor have public contact until the Wltested. elephant is tested unless approved by USDA.
Elephant TB Guidelines 17
prevent the emergence ofresistant strains of bacteria. For individuals exposed to TB (positive
skin test), but no signs of active disease (negative chest radiograph, negative sputum cultures),
treatment is typically with a single drug (INH).
The guidelines for the treatment ofTB in elephants are based on the assumption that animals
with known active disease are treated similarly to humans. However, for elephants, the
treatment period has been extended. For a category 3 elephant with negative cultures and
presumed exposure based on positive serologic response, i.e., positive ElephantTB STAT-PAK
(and MAPIA TM), treatment is a "modified" regime - with two drugs for 9 months. Skin testing
is not reliable in elephants . Acid-fast smears are not reliable on elephant trunk washes.
For humans, treatment of primary tuberculosis is to empirically administer 4 first line drugs
while waiting for antimicrobial sensitivity testing. This assures that initial treatment includes at
least 2 drugs to which the organism is susceptible . And, the additional number of antibiotics
results in more rapid clearance of bacteria from the sputum thereby decreasing the public health
risk.
Once susceptihility tests are received, and the sputum has reverted to being smear negative, the
number of drugs is decreased to two first line drugs for the remainder of treatment. When the
index case is known, and the index isolate is known to be susceptible to all anti~mycobacterial
drugs, then initial treatment may be limited to three drugs. However, in the va.st majority of
ca~es the index case is not lrnown with certainty and four drugs are given. Moreover, in regions
or situations when the frequency of resistance exceeds 10%, empiric initial therapy for humans
cons ists 0 f fi ve drugs .
The length of therapy for humans is currently 6 months for active tuberculosis. This includes the
initial period of 3-5 drugs as above and 2-drugs for the remainder of treatment. For individuals
with resistance to a single antibiotic, treatment is extended to 12 months with 2 drugs to which
the organism is susceptible. For individuals infected with multi-drug resistant tuberculosis
(MDR-TB), treatment is for at least 12 months with 2-4 drugs based on the susceptibility pattern
(lower numbers of agents are employed if the isolate is susceptible to INH or rifampin). Because
the long term outcome and efficacy of treatment for TB of non-human species is currently
unknown, treatment of elephants iE s1ructured for a 12-month course.
9. ANTI-TUBERCULOSIS DRUGS
Antituberculous agents are divided into first and second line agents. Fjrst line agents include
isoniazid, rifampin, pyrazinamide, ethambutol, and streptomycin. These are agents with the
greatest activity and the best side effect profiles . Second line agents include those with less
activity and/or greater side effects. Second line agents include capreomycin, ethionamide,
cycloserine, and thiacetazone. The fluoroquinolones (FQ; moxifloxacin, ciprofloxacin,
levofloxacin, and enrofloxacin) wlllie not considered as l't line agents have significant
bactericidal activity against M. tuberculosis. Moreover, published studies report the equivalency
of FQ substitution for ethambutol in the treatment of TB in humans and studies are underway to
investigate FQ use for the treatment of latent TB infection. Linezolid, a drug active against Gram
positive bacteria such as Staphylococcus aureus, MRSA, enterococcus, and VRE has also been
Elephant TB Guidelines 18
shown to have significant activity against M. tuberculosis and has been used successfully in
salvage regimens. Amikacin, an amino glycoside (as is streptomycin), is a mainstay in the
trea1ment of non-tuberculous mycobacterial infection and has been used in salvage regimens
against MDR-TB. Phannacoldnetic studies ofINH, RIP, EMB , and PZA in elephants have been
published (Maslow et aJ. 2005a, Maslow et a1. 2005 b, Zhu et a1. 2005, and Peloquin et al .
2006) .
Rifampin (RIF)
Mechanism of action: Rifarnpin is a semi synthetic derivative of rifamycin, an antibiotic
derived from the fungus Streptomyces mediterranei. Rifampin acts to inhibit the DNAdependent. RNA-polymerase thus blocking formation of messenger RNA (the first step in
Elephant TB Guidelines 19
protein synthesis).
Metabolism and excretion: Rifampin is acetylated in the liver. Both the unaltered and
acetylated drug is excreted into the bile. Rifampin is then reabsorbed whereas the acetylated
fonn is not.
Toxicity: The major toxicity of rifampin is hepatitis . Other side effects include gastrointestinal
upset, renal failure, hemolysis, acute renal failure, and thrombocytopenia. It is avoided in
pregnancy during the fIrst trimester because of possible teratogenicity.
Rifampin is also a strong inducer of the cytochrome P450 hepatic enzymes that may increase the
metabolism of concurrently administered drugs. A prime example is exogenously administered
steroids used for in vitro fertilization. For animals being treated for other conditions, potential
drug-drug interactions should be ruled out.
Pyrazinamide (PZA)
Mechanism of action: Pyrazinamide is a synthetic antibiotic derived from nicotinic acid. Its
mechanism of action is unknown; however the presence of an intact pyrazinamiciase is required.
Since Mycobacterium bovis lacks this enzyme, it is resistant to PZA.
Toxicity: Toxicities observed in humans include arthralgias and arthritis, hyperuricemia,
hepatitis, gastrointestinal upset, and photosensitivity (skin rashes).
Toxicity in elephants: The toxicity for elephants i~ unknown, however hepatitis may have been
observed. Similar adverse effects as documented for humans should be expected.
Route of administration: In humans, pyrazinamide is administered orally. rn elephants both
oral and rectal dosing have yielded acceptable blood levels. Pyrazinamide has been successfully
administered to bongo antelope via subcutaneous injection.
PZA is should not be given if M bovis infection is suspected since this organism is inherently
resistant to PZA."
Ethambutol (EMB)
Mechanism of action: Ethambutol is a specific inhibitor of the arabinosyl transferase thereby
inhibiting fonnation of arabinogalactose and lipoarabinomannan, which are the dominant lipids
Elephant TB Guidelines 20
Streptomycin
Mechanism of action: Streptomycin is an amino glycoside antibiotic derived from the fungus
Streptomyces griseus that acts on the 30S ribosome to inhibit protein synthesis.
Toxicity: Similar to other aminoglycosides, streptomycin admirUstration may result in auditoryvestibular and renal toxicity. Specific symptoms include ataxia, vertigo, nerve deafness, and
renal failure. Most symptoms are reversible if the drug is discontinued immediately after their
occurrence.
Toxicity in elephants: The toxicity for elephants is currently unknown but is likely the same as
for humans .
Elephant TB Guidelines 21
Amikacin
Mechanism of action: Amikacin is an amiooglycoside antibiotic that acts on the 30S ribosome
to inhibit protein synthesis. Isolates that are resistant to streptomycin may be susceptible to
amikacin.
Toxicity: Similar to other arninoglycosides amikacin administration may result in auditoryvestibular and renal toxicity. Specific symptoms include ataxia, vertigo, nerve deafness, and
renal failure. Most symptoms are reversible if the drug is discontinued immediately after their
occurrence.
Toxicity in elephants: The toxicity for elephants is currently unknown but is likely the same as
for humans.
Route of administration: Amikacin is administered via intravenous injection to humans.
Amikacin has been administered via intramuscular injection to bongo antelope yielding
acceptable serum levels (unpUblished). A pharmacokinetic study of amikacin in African
elephants has been conducted (Lodwick, LJ., Dubach., 1.M. and Phillips, L.G., 1994.
Pharmacokinetics of amikacin in African elephants. J Zoo Anim. Med 25: 367-375). There is
no published infoITJlatlon regarding amikacln in Asian elephants. Amikacin in one Asian
elephant given 1M 3 times a week at 14 mg/kg yielded good blood levels (acceptable levels in
elephants unknown) aod was eliminated ahnost completely from serum within 72 hours .
However, significant toxicity occurred with prolonged use of this drug at this dose (personal
communication, Dr. G Dumonceaux).
Other second line agents have not been used for mycobacterial infections in elephanis.
Clinicians contemplating the use of agents other than those listed should consult with the USDA
on an individual basis .
The fOUI first-line drugs used to treat tuberculosis in humans are isoniazid (INH), rifampin (RlF),
E1ephant TB Guidelines 22
pyrazinamide (PZA) and ethambutol (ErH). Second-line drugs used in cases of drug intolerance
or multi-drug resistant organisms include amikacin and a fluoroquinolone. Both
fluoroquinolones and linezolid have been used in cases of multidrug resistance in humans
(Veziris, N . et al. Fluoroquinolone-containing third-line regimen against Mycobacterium
tuberculosis in vivo. Antimicrob Agents Chemother 2003, 47: 3117 -22) .
Isoniazid
Rifampio
Pyrazinamide
Ethambutol
Dosage
(mglkg)
Route
5
4
4
10
30
30
Oral
Oral
Rectal
Oral only
Oral or rectal
Oral only
Formulation
premixed suspension
Powder
premixed suspension
Powder
Powder
Powder
Target
cone
(I-!g/ml)
3-5
3-5
3-5
8-24
20-60
2-5
Cmax
(hr)
1-2
0.5-1
0.25-0.5
2-4
l-2
1-2
The dosages quoted above are based primarily on the pharmacokinetic studies of drug
administration to the first herds of treated elephants as reported (Maslow et al 2005a., Maslow et
a12005b, Zhu et a12005, Peloquin et aI2006). Recent studies have demonstrated that lNH
achieves Cmax much more quickly than previously thought when administered rectally Dosages
are considered as estimates with the goal of achieving target serum concentrations as listed in
#10 below without causing significant side effects that interrupt treatment. Serum drug levels or
drug side effects may dictate that dosages be adjusted up or down accordingly. Sequential
MAPlA TM tests may also be used to monitor response to treatment (Lyashchenko 2006) . Second
line agents should only be considered and administered following consultation with the facility
USDA inspector.
Elephant TB Guidelines 23
line agents INH, RIP, PZA, and EMB (Maslow et aJ 2005a, Maslow et al 200Sb, Zhu et al 2005,
Peloquin et al 2006).
Blood levels should be determined to measure the maximal concentration of drug (Cmax). While
INR, PZA, and EMB are rapidly absorbed with a Cmax occuning between 1-2 hrs, drug
absorption may vary between elephants and may also vary drug to drug. Recent studies have
demonstrated that lNH achieves Cmax much more quickly than previously thought when
administered rectally. Importantly, the time to Cmax (Tmax) may vary over the course of
treatment due to multiple factors such as food intake, drug acceptance, etc. Thus, at the start of
treatment and periodically through the course of therapy it is important to measure drug levels at
multiple time points until Cmax for each drug and animal is determined.
For INH, PZA, and EMB it is recommended that drug levels be determined at lhr, 1.5hr, and 2
hr and for RIP at 2hr, 3hr, and 4hr except ifINH is administered rectally and then 15 min and)O
min blood levels are recommended to accurately measure the Cmax. Tfthe first measured time
point represents the greatest level for any drug, then Tmax may have already passed and earlier
time points should be assessed. Conversely, if the last measured time point represents the
greatest concentration for any drug, then Tmax may occur later than the range chosen and later
time points mould be assessed. During the initial phase of treatment, time ranges should always
be assessed to detennine the true Tmax .
NOTE: Target blood levels for anti-TB drugs in elephants have not been rigorously established.
Until further studies can be conducted, target blood levels of 80ti-TB drugs for elephants must
necessarily be based on human data. Although achieving blood levels comparable to humans is
the ideal goal, the attending veterinarian should be aware that there is unpublished evidence that
some elephants cannot tolerate anti-TB drugs at the doses required to achieve the above levels .
Isoniazid, in particular, has caused side effects. It may be necessary to reduce the dose of an
anti-TB drug to eliminate side effects, which may result in lower blood levels. The attending
veterinarian should carefully document observed side effects, dosage changes and associated
anti-TB drug levels in these cases. Variations to these Guidelines require consultation with the
facility USDA inspector.
www.elephanttag.ol1!;
www.eJephantcare.or g
Prior to any planned euthanasia of an elephant, trunk washes, blood for serology and any other
ancillary tests should be performed regardless of whether or not TB is suspected. In this way,
valuable data can be gathered to evaluate the efficacy of the current testing protocol. In the event
of a sudden death, collect post-mortem blood and separate serum for other tests.
Elephant TB Guidelines 24
It is recommended that a trained veterinary pathologist direct the necropsy if possible. 10 the
event of an elephant necropsy (elective or otherwise), contact Dr. Scott Terrell (Elephant SSP
Pathology Advisor) for further instructions and possible participation:
Scott P. Terrell, DVM, Diplomate ACVP, SSP Pathology Advisor, Disney's Animal Kingdom,
1200 N Savannah Circle, Bay Lake, FL 32830, W (407) 938-2746; H (407) 251-0545; Cell
(321 )229-9363; email Scott. P.Terrell@disney.com
The following infonnation is excerpted from the SSP Elephant Necropsy Protocol:
Elephant TB Guidelines 25
fixation (in buffered 10% formalin). In addition, search thoracic organs carefully for early stages
ofTB as follows: after removal of the lungs and trachea, locate the bronchial nodes at the
junction of the bronchi from the trachea. Use clean or sterile instruments to section the nodes.
Freeze half of the lymph node and submit for TB culture to NVSL or a laboratory experienced in
mycobacterial culture and identification (even if no lesions are evident). Submit sections in
formalin for histopathology. Carefully palpate the lobes of both lungs from the apices to the
caudal borders to detect any:firm B-B shot to nodular size lesions. Take NUMEROUS (5 or
more) sections of any suspicious lesions . Open the trachea and look for nodules or plaques and
process as above. Regional thoracic and tracheal lymph nodes should also be examined and
processed accordingly. Split the trunk from the tip to its insertion and take samples of any
plaques, nodules or suspicious areas for TB diagnosis as above. Look for and collect possible
extra-thoracic TB lesions, particularly if there is evidence of advanced pulmonary TB.
Any employee with a positive intradennal test (i.e. a positive intradermal reaction to purified
protein derivative (PPD) of M tuberculosis) should be evaluated for the possibility of active TB.
It is recommended that health care providers who manifest a positive PPD receive INH
prophylaxis unless there is a contraindication to treatment. Conversely, those declining
treatment are followed yearly with a chest radiograph and clinical evaluation to determine
whether they have developed active disease.
A positive skin test may result from either exposure to M tuberculosis, M. bovis, BeG injection,
or exposure to non-tuberculous strains of mycobacteria. The American Thoracic Society has
published guidelines for the interpretation of intradermal testing . If inoculation with BCG
occurred more than 10 years ago, a positive PPD test should not be considered a reaction due to
BeG, but should instead be considered as positive for exposure to TD.
Employees with acid-fast positive sputum smears should be removed from animal contact until it
is determined whether this represents infection with an organism of the M tu.berculosis camp lex
(M tuberculosis or M. bovis). Treatment guidelines and recommendations for contact with
animals and humans are available through state public health departments . At the present time
there is no known tramfer of non-tuberculous strains of mycobacteria between humans and
animals (or human to human) via aerosolization or any other route and thus, there are no
restrictions placed on animals or humans known to be colonized or infected such organisms.
Any facility housing a known culture-positive (M. tuberculosis complex) animal should develop
a program to protect employees from TB exposure, to include the use of appropriate face masks
(N95 HEPA filtered masks, certified by the National Institute for Occupational Safety and Health
to protect against IB), disinfection procedures, and the use of separate implements for infected
animals. The local public health department should be contacted for further guidelines.
Elephant TB Guidelines 26
Measures to protect staff from infected animals should include the use of respiratory (N95)
HEP A filtered masks during all direct or indirect contact with infected animals, such as cage
cleaning, medication administration, feeding, watering, etc. The facility should contact local
health agencies and should provide additional other protective gear such as gowns, gloves, etc.
No specific precautions are necessary for animals that are culture positive for mycobacteria other
than M. tuberculosis and M. bovis.
Best practices for the safe conduct of work in biomedical and clinical laboratories and animal
facilities in regards to Mycobacterium tuberculosis are listed in the 5th Edition of Biosafety in
Microbiological and Biomedical Laboratories published by the U.S. Department of Health and
Human Services in 2007. http://www .cdc.gov/od/ohsibiosfiv/bmbI5/BMBL 5th Edition .pdf
14. REPORTING
Tuberculosis is a reportable disease. Positive culture results must be reported to the State
Veterinarian and appropriate public health agencies.
15. APPENDICES
APPENDIX 1. REFERENCES CITED AND ADDITIONAL READING
Anon. 2003. Treatment of Tuberculosis. ATS, CDC and Infectious Diseases Society of America,
MMWR 52: No.RR-11 (June 20, 2003) 1-88 .
Auclair, B., Mikota, S.K., Peloquin, C.A., Aguilar, R., Maslow, J.N. 2002. Population
phannacokinetics of antituberculous drugs and treatment of Mycobacterium bovis infection in
bongo antelope (Tragelaphus eurycerus isaaci). J Zoo Wildl Med. Sep; 33(3): 193-203 .
Ball, R.L., Dumonceaux, G., Olsen, J.H., Burton, M.S., Lyashcbenko, K. Comparison oftnmk
wash results matched to mlJltiantigen print immunoassay (MAPlA TM) in a group of captive
Asian elephants (Elephas maximus). 2006. Proceedings American Association of Zoo
Veterinarians. 303 -304.
Centers for Disease Control and Prevention and National Institutes of Health. 2007. Biosafety
in Microbiological and Biomedical Laboratories, 5 th ed. u.s . Goverrunent Printing Office,
Washington, D.C. 143- t47~ http://www.cdc.gov/odJohs/biosfty/bmb15/BMBL 5th Ed ition.pdf
Davis, M. 2001. Mycobacten'um tuberculosis risk for elephant handlers and veterinarians. Appl
Occup Environ Hyg. 16: 350-353.
Greenwald, R., Lyashchenko, 0., Esfandiari, J., Miller, M., Mikota, S., Olsen, J.R., Balt, R.,
Dumonceaux, G., Schmitt, D., Moller, T., Payeur, J.B., Harris, B., Sofranko, D., and Waters,
W.R., Lyashchenko, K. 2009. Highly accurate antibody assays for early and rapid detection of
tuberculosis in African and Asian elephants. Clinical and Vaccine Immunology t6(5): 605-612.
Elephant TB Guidelines 27
Isaac, R. The elephant trunk wash - An update. Elephant Managers Association Annual
Conference. Orlando, Florida. November 9-11, 2001.
Isaza, R, and Ketz, C. A trunk wash technique for the diagnosis of tuberculosis in elephants.
Proceedings ofthe 39th international symposium of the European zoo and wildlife medicine.
Vienna, Austria . May 12-16, 1999. 121-124.
Lacasse, c., Terio, K., Kinsel, MJ., Farina, L.L., Travis, D .A., Greenwald, R., Lyashchenko,
K.P., Miller, M., and Gamble, K. 2007. Two cases of atypical mycobacteriosis caused by
Mycobacterium szulgai associated with mortality in captive African elephants (Loxodonta
a./ricana). J. Zoo Wildl. Med. 38 (1): 101107.
Landolfi, lA., Mikota, S.K., Chosy, 1., Lyashchenko, K.P., Giri, K., Gairhe, K., Teno, K.A.
2010 . Comparison of systemic cytokine levels in Mycobacterium spp. seropositive and
seronegative Asian elephants (Elephas maximus). J Zoo Wildl Med. 41(3): 445-455 .
Landolfi, LA., Schultz, S.A., Mikota, S.K., Teno, K.A. 2009. Development and validation of
cytokine quantitative, real-time RT-PCR assays for characterization of Asian elephant immune
responses. Vet Immunol Immunopathol. Sep 15; 131(1-2): 73-78.
Larsen, R.S., Salman, M.D., Mikota, S.K., Isaza, R., Montali, RJ. and Triantis, J. 2000 .
Evaluation of a multiple-antigen enzyme~linked immunosorbent assay (ELISA) for detection of
Mycobacterium tubercu.losis in captive elephants. J Zoo Wildl Med 31: 291-302.
Lewerin, S.S., Olsson, S.L., Eld, K., Roken, B., Ghebremichael, S., Koivula, T., Kallenius, G.,
and Bolske, G. 2005. Outbreak of Mycobacterium tuberculosis infection among captive Asian
elephants in a Swedish zoo . Vet Rec. 156(6): 171-175.
Lyashchenko, K., Singh, M., Colangeli, R.., and Gennaro, M.L. 2000. A multi-antigen print
Lrnmunoassay for the development of serological diagnosis of infectious disease. J ouma! of
Immunological Methods 242: 91-100.
Lyashchenko, K., Miller, M., and Waters, W.R. 2005. Application ofMAPJATM (Multiple
Antigen Print Immunoassay) and rapid lateral flow technology for tuberculosis testing of
elephants. Proe American Association of Zoo Veterinarians, 64-65.
Lyashchenko, KP., Greenwald, R., Esfandiari, J., Olsen, J.H., Ball, R., Dumonceaux, G.,
Dunker, F., Buckley, c., Richard, M., Murray, S., Payeur, J.B., Andersen, P., Pollock, J.M.,
Mikota, S., Miller, M., Sofranko, D., and Waters, W.R. 2006. Tuberculosis in elephants:
antibody responses to defined antigens of Mycobacterium tuberculosis, potential for early
diagnosis, and monitoring of treatment. Clin Vaccine Immunol 13(7): 722-732.
Maslow, J.N., Mikota, S.K., Zhu, M., Isaza, R., Peddie, L.R. , Dunker, F., Peddie, 1., Riddle, H,
and Peloquin, C .A. 2005. Population pharmacokinetics of isoruazid in the treatment of
Mycobacterium tuberculosis among Asian and African elephants (Elephas maximus and
Loxodonta africana). J Vet Pharmacal Ther. 28(1): 21-27.
Elephant TB Guidelines 28
Maslow, 1.N., Mikota, S.K., Zhu, M., Riddle, H., and Peloquin, C.A. 2005. Pharmacokinetics of
ethambutol (EMB) in elephants. J Vet Pharmacol Ther. 28: 321-323 .
Maslow, 1. Tuberculosis and other mycobacteria as zoonoses. 1997. Proceedings American
Association of Zoo Veterinarians. 110-115.
Micha.lak, K., Austin, c., Diesel, S., Bacon, J. M., Zimmennan, P., and Maslow, J. N. 1998.
Mycohacterium tuberculosis infection as a zoonotic disease: Transmission between humans and
elephants. Emerging Infect. Dis. 4: 283-287.
Mikota,S.K., Dumonceaux,G., Miller,M., Gairhe,K., Giri,K., Cheeran,J.V., Abraham,D.,
Lyashchenko,K., Larsen,S., Payeur,]., Waters,R., Kaufman,G. 2006. Tuberculosis in elephants:
An update on diagnosis and treatment; implications for control in range countries. Proceedings
International Elephant Conservation and Research Symposium, 109-118.
Mikota, S.K., Mi11er, M., Dumonceaux, G., Giri, K., Gairhe, K., Hamilton, K., Paudel, S.,
Vincent, B. Elephant tuberculosis diagnosis: implications for elephant management in Asian
range countries. 2006. Proceedings American Association of Zoo Veterinarians. 142-143.
Mikota, S.K., Peddie, L., Peddie, J., Isaza, R., Dunker, F., West, G., Lindsay, W., Larsen, RS.,
Salman, M.D., ChatteIjee, D., Payeur, J., Whipple, D., Thoen, c., Davis, S., Sedgwick, c.,
Montali, R.J., Ziccardi, M., and Maslow, 1. 2001. Epidemiology and diagnosis of
Mycobacterium tuberculosis in captive Asian elephants (Elephas maximus). J. Zoo Wildl. Med.
32: 1-16.
Mikota, S.K., Larsen, R.S., and Montali, R.I. 2000. Tuberculosis in elephants in North
American. Zoo Biology 19: 393 -403.
MoHer, 1., Roken, B.O., Lewerin, S.S., Lyashchenko, K., 2006 . The elephant Rapid Test (RT)
the future diagnostic test for TB (M. tuberculosis) in elephants? Call for a validation study in
Europe. Proceedings International Elephant Conservation and Research Symposium 119-124.
Moller,T., Roken,B., Petersson,L., Vitaud.,C., Lyashchenko,K.. 2005. Preliminary results of a
new serological test for detection ofTB-infection (Mycobacterium tuberculoSis) in elephants
(Elephas maximus and Loxodonta africanum) - Swedish Case studtes. Verh.ber.Erkrg .Zootiere.
42,173-181.
Montali, R.J., Mikota, S.K., and Cheng, LJ. 2001. Mycobacterium tuberculosis in zoo and
wild1ife species. Revue Scientifique et Technique Office International des Epizooties 20(1):
291-303.
Montaii, R.J., Spelman, L.H., Cambre, RC., Chatterjee, D. and Mikota, S.K. Factors influencing
interpretation of indirect testing methods for tubercuJosis in elephants . 1999. Proc . Amer.
Assoc . Zoo Vet. 109-112.
Murphree, R., Dunn , J .R, Warkentin, I.V., SChaffner, W., and Jones, 1.F. 2010 . Mycobacterium
tuberculosis infection among employees of an elephant refuge. 2010 National Tuberculosis
Elephant TB Guidelines 29
Elephant TB Guidelines 30
APPENDIX 2. ACKNOWLEDGMENTS
The following individuals have contributed to the historical development of these
GuideUnes:
Dr. Wilbur Amand, Director Emeritus American Association of Zoo Veterinarians
Dr. Miava Binkley, USDA, Animal Care
Dr. Genevieve Dumonceaux, Florida Aquarium
Dr. Freeland Dunker, Steinhart Aquarium
Dr. Murray Fowler, University of California., Davis
Dr. Werner Heuschele, San Diego Zoo (in memoriuro)
Dr. R.amiro Isaza, University of Florida - Gainesville
Dr. Barbara Kahn, USDA, APHIS, Animal Care
Dr. Scott Larsen, University of California, Davis
Dr. William A. Lindsey, Feld Inc.
Dr. Konstantin Lyashchenko, Chembio Diagnostic Systems, Inc.
Dr. Joel Maslow, University of Pennsylvania
Dr. Bob Meyer, USDA, APHIS, Veterinary Services
Dr. Susan K. Mikota, Elephant Care International
Dr. Richard Montali,
Dr. C. Douglas Page, Jacksonville Zoo
Dr. Linda Peddie and Dr . James Peddie, America's Teaching Zoo, Moorpark College
Dr. Mo Salman, Colorado State University
Dr. Dennis Schmitt., Feld Inc.
Dr. Scott Terrell. Disney' s Animal Programs
Dr. Dominic Travis, Lincoln Park Zoo
Dr. Charles Thoen, Iowa State University
Dr. Gary West, San Antonio Zoo
Ms. Diana Whipple, USDA, ARS, National Animal Disease Center
Dr. Michael Ziccardi, University of California, Davis
The following individuals are members of the U.S .Animal Health Association TB Scientific
Su bcommittee:
Dr. Chuck Massengill, Missouri Department of Agriculture
Dr. Susan K. Mikota, Elephant Care International
Dr. Michele Miller, Palm Beach Zoo
Dr. Kathy Orloski, USDA, APillS, Veterinary Services
Dr. Janet B . Payeur, USDA, APHIS, National Veterinary Services Laboratories
Dr. W. Ray Waters, USDA, ARS, National Animal Disease Center
Elephant TB Guidelines 31
Summary
A trunk wash is a practical method of collecting a sample from an elephant's distal respiratory
tract for Mycobacterium culture and is the technique recommended in the "Guidelines for the
Control of Tuberculosis in Elephants" by the National Tuberculosis Working Group for Zoo and
Wildlife Species. The procedure, however, is potentially dangerous to the handlers and requires
cooperation of the elephant. Because of the limitations of using culture results as a screening test,
the trunk wash results should be interpreted with care. A positive culture result identifies an
elephant that is shedding tuberculosis organisms whereas a negative result is non-diagnostic.
Introduction
Tuberculosis in Asian elephants (Elephas maximus) has been sporadically reported in the literature
for many years (1, 2). The isolation of Mycobacterium tuberculosis from elephants in the United
States has resulted in the development of the "Guidelines for the Control of Tuberculosis in
Elephants" by the National Tuberculosis Working Group for Zoo and Wildlife Species
(http://www .aphis. usda. govfacIE I ephTBGuidelines2000. htrnl). Compliance wi th this po Iicy
requires that all elephants have annual mycobacterial cultures. In these guidelines, the trunk wash
is recommended as the most practical method of obtaining a culture sample from an elephant. This
paper describes the trunk wash technique as the authors are currently using it.
Procedure
A routine screening of an elephant should consist of a series of three trunk wash samples collected
on separate days withjn a one-week period. Trunk washings should be collected in the morning
and prior to water being offered to the animal. These recommendations are made in an attempt to
obtain a representative sample of the nasal flora from the previous night, and to avoid the dilution
effect caused by elephants drinking water with their trunks.
Elephant TB Guidelines 32
The elephant's trunk is manually restrained by the handlers so that the tip is held up. The 60 ml
syringe filled with sterile saline is then inserted into one of the nostrils and the saline quickly
flushed into the trunk. The handler then lifts the trunk tip as high as possible to help the fluid flow
as far into the trunk as possible. The I gallon plastic bag is then slipped over the trunk tip and the
tip of the trunk is lowered to allow the fluid to drain. If possible, the elephant is anowed to exhale
into the bag during this collection phase of the procedure. A good sample should retrieve a
significant portion of the saline that was placed into the trunk (about 40 rnl). The sample should
contain visible mucus from the inside of the trunk and often contains dirt and food particles that
are nonnally found inside the trunk . The collection of moderate amounts of foreign material does
not invalidate the sample. If, however, the collector feels the contamination is excessive, a second
flush may be attempted.
Once the sample is collected in the plastic bag, it is carefully transferred into a labeled container.
Ideally, the sample is refrigerated and sent directly to a laboratory for processing and
mycobacterial culture. If the sample cannot be sent directly for culturing, it may be frozen in a
regular freezer (-20 to -10 0c) until it can be sent to the laboratory. Often the recommended three
daily cultures samples are collected and frozen until all samples are collected and the batch of
samples can be sent to the laboratory together.
Discussion
Identification of a M. tuberculosis infected animal has significant management implications to
both the animal and the collection. Management of the infected animal may require isolation of
the exposed herd, potential removal of the animal from exhibit or shows, and if elected, treatment
of the animals and exposed herd which can be very expensive. In the worst case, a positive
diagnosis may lead to euthanasia of the infected animals. For these reasons , the screening test
selected needs to be definitive and have as few false positives as possible. A positive culture of M.
tuberculosis is, therefore, the only diagnostic test result used as a basis for making decis ions in the
guidelines.
The trunk wash as a method of collecting a culture sample from elephants was selected by the
National Tuberculosis Working Group for Zoo and Wildlife Species because it is a practical
method of obtaining a culture sample from a large proportion of the elephant population. The
procedure requires no sedation or undue stress to the animal. Additionally, the procedure requires
no specialized or expensive equipment.
An important consideration of this procedure is that it can potentially be very dangerous to the
handlers. This is particularly true when attempted on an uncooperative elephant, because any
attempts to manually restrain the trunk in an uncooperative elephant can lead to injury. The time
spent training the elephant to accept this method will greatly increase the efficiency and safety of
the procedure. In some cases, with potentially dangerous or unpredictable animals, an increased
level of handler safety can be obtained by having the animal lie in sternal or lateral recumbency
prior to sample collection. This technique does not guarantee safety or successful sample
collection, as it still requires cooperation of the animal and does not replace adequate training. In
the case of elephants managed under protective contact, the anima l's trunk can be handled though
a set of bars. This method still requires that the animal is fully cooperative and, therefore, usually
Elephant TB Guidelines 33
References
1. Mikota, S., Sargent, E.L., Ranglack., G.S. Medical Management of Elephants. West
Bloomfield, MI, Indira Publishing House. 1994.
2 . Mikota, S.K., Peddie., L., Peddie, J., Isaza, R., Dunker, F., West, G ., Lindsay, W., Larsen,
R.S., Salman, M.D., ChatteIjee, D., Payeur, J., Whipple, D ., Thoen, c., Davis, D.S ., Sedgwick,
c., Montali, R.J., Ziccardi, M., and Maslow, I. Epidemiology and diagnosis of Mycobacterium
tuberculosis in captive Asian elephants (Elephas maximus). J Zoo Wild) Med 2001 Mar. 32 (1):
1-16.
3. Michalak, K., Austin, C., Diesel, S., Bacon, I .M., Zimmerman, P., and Maslow, J.N.
Mycobacterium tuberculosis infection as a zoonotic disease: Transmission between humans and
elephants. Emerg Infect Dis. 1998. Apr-Jun 4(2): 283-287.
Elephant TB Guidelines 34
Send trunk washes to NVSL either frozen or on icepacks by overnight express (Federal
E;q>ress handles diagnostic samples). Containers should be leak proof and double-bagged (50 ml
conical screw-top centrifuge tubes are preferred) and are available free of charge from NVSL.
If lesions are submit1ed for culture, tissues should be frozen and sent on ice packs
-overnight. Lesioned tissues should be split and Yl should be sent to the histopathology lab so
peR can be run to see if the tissne is compatible for tuberculosis. There is no charge fOT
histopathology on lesioned tissue.
Use the VS Form 10-4 for submission, not the VS 6-35 form found in the TB kit. If the
forma1ized tissue is sent separately from the frozen tissue, please indicate on the submission
forms that there are 2 separate packages coming from the same animal so that the reports
can be combined and accession numbers coordinated when they reach NVSL. It is also
helpful to call or email NVSL contacts when sending TB suspects to schedule testing and relay
any relevant history of the case.
NVSL Trunk wash cost: $98 per sample for processing which includes a Oen Probe DNA
probe on any isolate. If the sample is positive for mycobacteria and speciation is requested, the
charge is $122.00 per sample which includes biochemical analysis, 165 rDNA sequencing
analysis, spoliotyping and VNTR genotyping. DNA fingerprinting of M tuberculosis or M bovis
isolates is also available. Antimicrobial susceptibility testing is available for M. tuberculosis
Elephant TB Guidelines 35
complex organisms for $112.00 per isolate. Please contact NVSL at (SI5) 337-7388 for test
schedule.
To establish an account at NVSL for billing, contact Connie Osmundson (515) 337-7571
Email: Coonie.J.Osmundson@aphis.usda.gov .
Of
(User fees as of October 1, 2010). Call lab before shipping samples for current prices and
schedule of testing or check prices at the NVSL web site:
http://www.aghis.usda.goY/animal health/lab info serv ices/diagnos tests.sht1l11
Mycobacteriology Lahoratory at National Jewish Medical aod Research Center
National Jewish Medical and Research Center
Director: Leonid H eifets. M.D.
1400 Jackson St.
Denver, CO 80206
(303) 398-1384
E-mail: heifets l@nic.org
For price list, shipping instructions, and requisition form:
hnp: //www.national jewish.org/research/clinical-labs/aboutlleam/mvcobac/index.aspx
Serum sample submission: it is important to protect the samples from light by wrapping the tubes
in tinfoil and to separate the serum and freeze it without delay, transferring the serum to a tube or
cryovial that is also wrapped in tin foil. Samples should be sent on dry ice as well.
mSTOPATHOLOGY
Scott P. Terrell, DVM, Diplomate ACVP
SSP Pathology Advisor
Disney's Animal Kingdom
1200 N Savannah Circle
Bay Lake, FL 32830
W (407) 938-2746;
H (407) 251-0545;
Cell (321) 229-9363;
Email Scott.P.TerreJI@disney.CDm
Send sections in formalin of any gross lesion and complete set of tissues including lung, liver,
spleen, mesenteric lymph nodes, bronchial lymph nodes and other major organs. Use leak proof
container.
Elephant TB Guidelines 36
Elephant TB Guidelines 37
Elephant TB Guidelines 38
1.
Purpose
The purpose of this Standard Operating Procedure (SOP) is to describe the procedure for
processing elephant trunk washes for the isolation of Mycobacterium tuberculosis used in the
Mycobacteria and Brucella (ME) section.
Materials
2.1
2.2
2.3
2.4
2.5
2.6
2.7
Elephant TB Guidelines 39
2.8
2.9
2.10
2.11
2.12
2.13
2.14
3.
sterilize solution thru a O.22)lm filter into sterile jug with bell end.
Dispense (wearing gloves), 20 mJ into a 50 ml sterile conical tube.
Caution - chemical is carcinogenic.
10 ml
2 .7.4 Vancomycin (NVSL #40151)
2.7 .4.1 Combine 9.346 gm with 1000 ml of distilled Super Q H20 and mix
weI\. Filter sterilize. Dispense into 50 rol sterile tubes in 20 .5 ml
amounts.
5 m1
2.7.5 Amphotericin B (NVSL #40154)
2.7.5.1 Add 10 ml wann sterile distilled H20 to a 100 mg Amphotericin B
(Fungizooe) . Shake gently until dissolved and dispense as
requested.
2.7.6 Combine BHl broth and Super QfuO . Autoclave for 20 min at 121C.
Cool to 50C. Remove 25 rol of cooled broth and discard. Add Nalidixic
Acid, Vancomycin, and Amphotericin B. Mix wel1. D ispense in tubes
and cover with foil became the Amphotericin B is light sensitive . Store in
-20C freezer. Media is good for 3 months.
3 7C CO~ incubator preferred
Media set-up (one tube of each per sample):
2.9 .1 Middlebrook 7HI0 w/glycerol; NVSL #10941 or BBL Middlebrook and
Cohn 7HIO Agar tubes (BBL #220959).
2.9.2 Middlebrook mIl w/glycerol; NVSL #10942 or BBL Seven H11 Agar
tubes (BBL #221392) or BBL Selective Seven H11 Agar tube (BBL
#297639).
2.9.3 Stonebrinks; NVSL #10451
2.9.4 Mycobactosel L-J medium (BBL #221414)
2.9.5 Bactec l2B medium vial with Panta and Erythromycin (32 ,ug/ml)
I ml tuberculin syringes
5 ml syringes
Slant trays, media tube baskets
Vortex
Sterile swabs
Procedures
3.1
3.2
Pour 10 to 12 ml (or whatever is left if < 10 ml) of the remaining trunk wash into
a 15 m1 conical centrifuge tube for storage.
3.2.1 These 15 ml centrifuge tubes are stored at -20 2 0 C for a minimum of 8
weeks or until the bacterial isolation procedure is completed.
3.2.2 Samples from cases in which no isolation was made are retained in a
-20 2 C freezer for at least 6 months .
Elephant TB Guidelines 40
3.3
3.4
3.5
3.6
3.7
3.8
3.2.2.1 Samples that have no isolation and are older than 6 months can be
discarded by the procedure in the current version ofMBSOP0008 .
3.2.3 Samples from cases in which mycobacteria have been isolated wiU be
retained for one year and stored in a -20 0 C freezer.
Allow the 10 - 12 ml of the trunk wash in the 50 ml conical tube to stand
Or
undisturbed for 15 - 20 minutes to allow sediment to settle to the bottom.
Alternately:
Pulse spin the 10 - 12 ml of trunk wash in the 50 ml centrifuge to spin down
excess sediment. This can be accomplished by centrifuging for 1 minute and 40
seconds at 3000RCF, lOoe, using the Sorvall RC 3BP centrifuge.
Slowly pour the supernate, trying not to disturb the sediment, into a sterile 50 rnl
conical centrifuge tube.
Prepare the N-Acetyl-L-cysteine (NALC)lNaOH- sodium citrate solution
according to the following proportions:
Volume (ml)
NaOH- sodium citrate a (mn
NALC (g)
50
0.25
50
100
0.50
100
150
0.75
150
200
200
1.00
300
300
1.50
a. 1: 1 mixture of 4% NaOH to 2.9% sodium citrate
b. Allow NALC to dissolve in solution before use
c. Discard this solution after 24 hours
Add an equal amount of the NALC solution to the trunk wash supernate up to a
maximum of 10 ml using a sterile pipette .
3.6.1 Be careful to avoid cross contamination of samples when adding the
NALC solution.
Vortex or vigorously hand shake for 20 5 seconds
Routinely allow the trunk wash to remain in contact with the NALC for 15 5
minutes.
3.8.1 If the sample is extremely cloudy or appears to be contaminated, the
NALC solution may need to remain in contact with the sample for up to
20 5 minutes.
NALC is a mucolytic agent and this procedure reduces or eliminates
contaminating bacteria while releasing Mycobacteria which may be
trapped in mucin and cells, allowing them to grow. Observe the tube for
clearing before proceeding to the next step.
Add enough sterile distilled water to the NALC/wash solution to fill the
centrifuge tube
Centrifuge the waterlNALC/wash solution for 20 minutes at 6000g and lOoe.
Carefully pour off and discard the supernatant.
Re-suspend the sediment with 2 mls of sterile distilled water and 1 ml of the
Johne 's antibiotic mixture.
3.8.2
3.9
3.10
3.11
3. 12
Elephant TB Guidelines 41
3.13
3.14
3.15
3.16
3.17
3.18
3.19
3.20
3.20.1.3
Elephant TB Guidelines 42
3.20.2.3
4.
References
4 .1
Della-Latta, P. and 1. Weitzman. 1998. Mycobacteriology. p. 169-204 In H.D.
Isenberg (ed .), Essential Procedures/or Clinical Microbiology. American
Society for Microbiology, Washington, D .C.
4.2
Zimbro, MJ . and O.A. Power. 2003. Difco & BBL Manual- Manual of
Microbiological Culture Media. Becton, Dickinson and Company, Sparks, 1vID.
Elephant TB Guidelines 43
Elephant TB Guidelines 44
INTERNET
These guidelines are available on the Internet at the following sites:
Elephant TB Guidelines 45
-------------------------------------
Submitter:
Address:
-----------------------------------------------
Tel:
Animal Information
AsiAn [)
African [) ISIS#
Name
Sex: [ J male
Studbook # _--,------:-_ _
Age: _______ [ 1actual [ J estimate
r J female
Ship samples overnigbt frozen witb sbipping box marked "PLACE 1N FREEZER UPON ARRIVAL"
Elephant TB Guidelines 46
1ll1derstand that all results and recorrunendations regarding the individual elephant will be kept
confidential .
_ _ Yes,
results.
_ _
No,
r agree to allow the SSP to use our sample for designated research and testing
r do Dot consent to the use of our sample and test results unless specified.
Signature, title
Printed name
Institution
Date
Phone number
Email address
Address
Conunents: ______________________________________________________
Elephant TB Guidelines 47
Group 1: Culture
nega tive; S AT-PAK
DOD-reactivc:; 80 exp08Bre
past lllDOBtbJ
No treatment or travel
restr'ctions
I
If exposed to untested
elephant in previous 3
months repeal STAT-PAK
in 3 months
TB Management Group 2
G oup 2: Culture
aeptive; STAT-PAK
DOD-reactive; exposed t~
cliiture positive an Imal in
palt 12 mOD hs
and STAT-PAK@nonreactive
Elephant TB Guidelines 48
TB Management Group 3A
No treatment or tray I
restrictions
TB Management Group 3B
I
Cultur q 3 months 1st y~ar: q 6
months ne. t 2. Repeat
MAPIAl'MIDPP q 6 months
Ist 3 years. Resume annual
testing if cultures negative and
MAPIAIDPP non-reactive
after 3 years
1
No travel or public contact 1st
year; restrictions removed if
results unchanged. If results
change, elephant will change
group.
Elephant TB Guidelines 49
TB Management Group 3C
Elephant TB Guidelines 50
Management Group 3D
Elephant TB Guidelines 51
Management Group 4
Group 4: Culture
positive f()r
M. tuberculnm
CoalpleX
I
Perfonn STAT-PAK and
Initiate tre&tmet1[
efer to text
Maintain in
pemlanent qUM8Jttine
- refer to text
\
No travel or public
contact until
No travel or pub-lie
contact until
treatment
treatment is
completed
according to these
completed
according to these
Guidelines
Guidelines.
Euthanasia - refer to
text